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05 March 2011, Volume 15 Issue 10 Previous Issue    Next Issue

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Bone marrow mesenchymal stem cells isolated from rats by whole bone marrow adherent culture in vitro and PKH26 labeling Ye Xiao-feng, He Yuan-Li, Wang Xue-feng, Fu Xia-fei 2011, 15 (10):  1711-1714.  doi: 10.3969/j.issn.1673-8225.2011.10.001 Abstract ( 283 )   PDF (1188KB) ( 446 )   Save BACKGROUND: In vitro culturing, amplification and labeling are important links to harvest labeled rat bone marrow mesenchymal stem cells (BMSCs) that are satisfactory to animal experiment. OBJECTIVE: To explore a convenient and practical method for separating and labeling BMSCs using whole bone marrow adherent culture, and labeling with PKH26 in vitro. METHODS: BMSCs were isolated and cultivated from the bone marrow of rats by whole bone marrow adherent culture. Further purification was achieved by expansion at serial passages. The passage 3 BMSCs were cultured and labeled with PKH26. The growth, fluorescence intensity and serial subcuhivation of labeled BMSCs were analyzed with fluorescence microscope. The proliferation ability of these labeled cells was tested by MTT. RESULTS AND CONCLUSION: BMSCs were isolated and purified successfully and effectively by the method of whole bone marrow adherent culture. The labeled BMSCs appeared red fluorescence. After 3 passages of serial subcultivation, the fluorescence intensity and the labeling rate of BMSCs were gradually decreased. The biological features such as morphology, growth in vitro were not affected by labeling. BMSCs can be successfully cultivated by whole bone marrow adherence method conveniently. Labeling the BMSCs with PKH26 is an effective and practical method. Related Articles | Metrics

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Effects of high-frequency pulsed electromagnetic fields on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells Cui Xiang-rong, Su Wei, Huang Zhao, Qin Wan-an 2011, 15 (10):  1715-1720.  doi: 10.3969/j.issn.1673-8225.2011.10.002 Abstract ( 299 )   PDF (2206KB) ( 460 )   Save BACKGROUND: The researches about low-frequency pulsed electromagnetic fields (PEMFs) interfering with bone marrow mesenchymal stem cells (BMSCs) proliferation and differentiation are many, but there is no report concerning high-frequency (> 300 MHz) PEMFs. OBJECTIVE: To explore whether high-frequency (> 300 MHz) PEMFs can promote proliferation and osteogenic differentiation of BMSCs. METHODS: BMSCs were isolated from Sprague-Dawley rats. The third passage of BMSCs were obtained and divided into four groups: osteogenic induction group, osteogenic induction+PEMF irradiation group, PEMF irradiation group and blank control group. Changes in morphology, number and amount of total protein were observed during BMSC culture in each group. RESULTS AND CONCLUSION: The number of BMSC bodies was increased, but the differentiation ability was weak following PEMF irradiation. The speed of cell proliferation and content of total protein were lower in the PEMF irradiation group compared with blank control group (P < 0.05). However, cell apoptotic rate was greater in the PEMF irradiation group compared with blank control group (P < 0.05). Results indicated that promoting effects of high-frequency PEMFs on osteogenic differentiation of BMSCs are not obvious. High-frequency PEMFs can suppress BMSC proliferation and contribute to BMSC apoptosis. Related Articles | Metrics

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Culture and identification of rat bone marrow mesenchymal stem cells Li Xiao-feng, Zhao Jin-min, Su Wei, Cui Xiang-rong, Luo Shi-xing, Ma Ai-guo 2011, 15 (10):  1721-1725.  doi: 10.3969/j.issn.1673-8225.2011.10.003 Abstract ( 726 )   PDF (1720KB) ( 2001 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are not rich in bone marrow, and are difficult to culture in vitro. It is crucial to tissue engineering and to the cell experiments in vivo or in vitro that isolating and culturing in vitro BMSCs with high purity, strong vitality and homogeneous biological characteristics. OBJECTIVE: To establish a method of isolation, cultivation and purification of rat BMSCs in vitro, to observe cell morphology, and to assess surface markers and multi-directional differentiation capacity. METHODS: BMSCs from rats were isolated, cultured and purified by the whole bone marrow adherence method, for morphology observations, the growth curve was drawn, cell cycle was analyzed, cell surface markers were assessed by flow cytometry, and BMSCs were induced to differentiate into osteoblasts and adipocytes. RESULTS AND CONCLUSION: BMSCs from rats were spindle cell-based, showing radial colony arrangement. Cells kept strong growth and could passage in continuous and stable manner over 10 passages. The growth curve and cell cycle demonstrated that BMSCs were consistent with the growth characteristics and good activity of normal cells. At the third passage, BMSCs were negative for CD34 and CD45, but positive for CD44, CD90 and CD105. Following induction, oil red O staining, alkaline phosphatase staining, von Kossa staining and alizarin red staining produced a strong reaction in cells. Whole bone marrow adherence method is simple and can isolate, purify and amplify BMSCs in vitro. The obtained cells have general biological characteristics of mesenchymal stem cells, and also have potentiality of multi-directional differentiation. This experimental method has important practical significance to provide adequate source of seed cells for tissue engineering. Related Articles | Metrics

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Effects of myocardium lysate on the differentiation of rat bone marrow-derived mesenchymal stem cells into cardiac myocyte in vitro Li Qin, Zhao Wen-jing, Kou Ya-li, Hu Li-xia, Chen Wei-ping 2011, 15 (10):  1726-1730.  doi: 10.3969/j.issn.1673-8225.2011.10.004 Abstract ( 377 )   PDF (1610KB) ( 449 )   Save BACKGROUND: Stem cells have the characteristics of “differentiation dependent on enviroment”, little research have focused on myocardium lysate that simulating the microenvironment of cardiocyte induce bone marrow mesenchymal stem cells (BMMSCs) differentiated into cardiocyte-like cells. OBJECTIVE: To investigate the effects of myocardium lysate on the differentiaion of bone marrow mesenchymal stem cells into myocardial-like cell. METHODS: Changes of morphological characteristics of the second passaged BMMSCs that cultured with medium contained myocardium lysate for 5 weeks were observed. After the procedure of induction, α-Actin was used to observe whether BMMSCs differentiate into cardiocyte-like cells, hematoxylin-eosin staining was used to identify whether intercalated disk was existed, while electron microscopys was used to assess the ultrstructure of the induced cells. RESULTS AND CONCLUSION: Most of the induced cells like bamboo in microscopys, positive reaction after α-actin immunocytochemical stain showed that the BMMSCs were differentiated into myo-like cells. Intercalated disk, well sequenced sarcomeres and myofilaments of the cytoplasm observed revealed the bamboo-like cells have the typical morphology of cardiocyte. Spontaneous beating myotube-like structures generated at 12 days after induction. In brief, the myocardium lysate can imitating the microenvironment of cardiocyte, induce BMMSCs differentiated into cardiocyte-like cell which have the typical morphology of cardiocyte and the corresponding functional activities. Related Articles | Metrics

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Effect of aging on proliferative capacity of human bone marrow mesenchymal stem cells in vitro Wang Xiao-yan, Wang Zhi-wei, Wang Qing-shan, Xie Dong-fang, Chen Bo-hua 2011, 15 (10):  1731-1735.  doi: 10.3969/j.issn.1673-8225.2011.10.005 Abstract ( 301 )   PDF (1341KB) ( 361 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) from human beings can proliferate and differentiate to repair the damages of tissues and organs. The reparation capacity of tissues and organs decreases with age. Question arises if the proliferation capacity of MSCs deceases with age. OBJECTIVE: To evaluate the effect of aging on the proliferative capacity of human MSCs in vitro. METHODS: Fifteen bone marrow specimens were obtained from the patients in Qingdao Central Hospital, who received the operation of bone because of bone fracture or hip diseases. They were divided into three groups: 5 children aged from 7 to 12 years; 5 adults aged from 20 to 55 years; 5 elder aged from 60 to 90 years. The bone marrow isolates were taken out from patients during operation. The mononuclear cells were isolated from bone marrow using density gradient centrifugation and seeded into the culture flask. On the fifth day, colony forming units were counted under microscope. When the cells reached a confluence, they were passaged and further culture was preceded in two ways: 192 cells were seeded in two 96-well plates to study the cloning efficiency of single cells; 12 500 cells were seeded in T25 flask at the density of 5×103/cm to study the proliferation rate of cells. RESULTS AND CONCLUSION: There were no correlation between colony forming units and proliferation rate of MSCs cultured in vitro and the age (P > 0.05), but the cloning efficiency of single cells decreased with age (P < 0.01). The number and proliferation capacity of MSCs do not decrease with age, but the uncommitted stem cells that could proliferate to single cell clones do deplete with age. Related Articles | Metrics

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Hydrogen sulfide protects human bone marrow mesesenchymal stem cells against cisplatin-induced damage Li Jing-chun, Huang Gang, Yong Bi-cheng, Xu Ming-hong, Wang Yao-fei, Zhang Long-juan 2011, 15 (10):  1736-1740.  doi: 10.3969/j.issn.1673-8225.2011.10.006 Abstract ( 324 )   PDF (1372KB) ( 429 )   Save BACKGROUND: Due to serious myelosuppression caused by hemopoietic stem cell damage when using cisplatin chemo-treatment, there is no effective cytoprotective agent of chemo-treatment to lessen its adverse reaction. OBJECTIVE: To explore the protection of hydrogen sulfide (H2S) against cisplatin–induced human bone marrow mesesenchymal stem cells (hBMSCs) damage. METHODS: Sodium hydrosulfide (NaHS) as a H2S donor. hBMSCs treated with different concentrations of sodium hydrosulfide and cisplatin for 48 hours, the cell survival rate was measured by MTT assay; the morphological change of apoptotic cells was tested by using the chromatin dye Hoechst 33258; the proteins expression were detected with Western blot techniques. RESULTS AND CONCLUSION: MTT detection showed that cisplatin chould reduce the survival rate of BMSCs, and NaHS dose-dependently blocked the inhibition of hBMSCs cells growth induced by cisplatin. When hBMSCs cells were treated by both NaHS (0.5, 1 mmol/L) and cisplatin (20 mg/L) for 48 hours, the NF-κB (p65) was up-regulated, but the IκB-α was down-regulated. When hBMSCs cells were treated by both NaHS (1 mmol/L) and cisplatin (20 mg/L) for 6 hours, the level of phosphorylated protein kinase 1/2 was significantly ascended. The results showed that H2S chould protect hBMSCs against cisplatin-induced damage, which may be associated with the activation of protein kinase-NF-κB signaling pathway. Related Articles | Metrics

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Recombinant human bone morphogenetic protein-2 combined with basic fibroblast growth factor promotes vascular endothelial growth factor expression and osteogenic potential of bone marrow stromal cells in rabbits  Chang Qi, Huang Chang-lin, Huang Tao 2011, 15 (10):  1741-1744.  doi: 10.3969/j.issn.1673-8225.2011.10.007 Abstract ( 410 )   PDF (1192KB) ( 333 )   Save BACKGROUND: Whether recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF) can reach the dual role of the strength of osteogenesis and promotion of bFGF expression is need to know in vitro or in vivo experiment. OBJECTIVE: To investigate the effects of rhBMP-2 and bFGF on the vascular endothelial growth factor (VEGF) expression and osteogenic potential of marrow stromal cells (MSCs). METHODS: The MSCs of the femur of the rabbit were procured and cultured by MSCs cultured in vitro. The samples were treated with rhBMP-2 or (and) bFGF. After culturing for five days, the morphology of cells, proliferation, alkaline phosphatase activities (APA), bone clusters, and the ratio of VEGF-positive cells were detected. RESULTS AND CONCLUSION: The effects of combined use of BMP-2 and bFGF on cell counting, APA, percentage of mineralized area, and the ratio of VEGF-positive cells were better than that of using rhBMP-2 or bFGF singly. The results showed that a rational combined use of rhBMP-2 and bFGF can not only enhances multiplication ability and osteogenic transformation of MSCs, but also promotes VEGF expression of the proliferation of VEGF. Related Articles | Metrics

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Human umbilical cord mesenchymal stem cells differentiate into Schwann cells in vitro Wang Guang-yu, Zhao Fang, Hou Rong-rong, Zhu Lü-yun, Ma Li-cheng, Hu Li-ye, Li Xiao-ling, Yang Shao-ling, Shan Wei 2011, 15 (10):  1745-1749.  doi: 10.3969/j.issn.1673-8225.2011.10.008 Abstract ( 316 )   PDF (1229KB) ( 396 )   Save BACKGROUND: Human umbilical cord Wharton's Jelly mesenchymal stem cells avoid the ethical constraints, which have rich resources, and can be used as seed cells for tissue repair. OBJECTIVE: To observe the feasibility of umbilical cord Wharton's Jelly mesenchymal stem cells (UC-MSCs) differentiated into Schwann cells in vitro. METHODS: UC-MSCs were isolated and cultivated. The surface markers of UC-MSCs were identified by flow cytometry. UC-MSCs were induced and differentiated into Schwann cells by two stages with neurons medium, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), retinoic acid (RA), platelet-derived growth factor (PDGF). Morphologic changes of cells were observed under inverted microscope. The expression of Nestin, S-100, and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical stain method. Specific proteinum expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot technique. RESULTS AND CONCLUSION: Morphous of UC-MSCs were changed at 7 days culture. A part of UC-MSCs changed into fusiform shape. The primary cell could achieve 80%–90% confluence about 10 days, the morphous of UC-MSCs were fusiform shape. Cells isolated from human umbilical cord expressed specific mesenchymal stem cells surface markers: CD44 (91.4%), CD29 (91.3%), CD105 (99.2%); without expression CD34 (0.2%), CD45 (0.9%), CD14 (0.6%). Morphous of UC-MSCs changed from short fusiform shape to long fusiform shape or fusiform, cells assembled and some global cell groups formed after the first induction stage. After the second induction stage, some long fusiform shape cells grown from global cell groups. Morphous of major cells were long fusiform shape after 96 hours with the phenomenon of multipolar. The immunocytochemical stain showed that long fusiform cells have Schwann cells specific GFAP and S100 protein staining. The results showed that UC-MSCs could be differentiated into Schwann cells in vitro. Related Articles | Metrics

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Biological activity and restriction of human umbilical cord mesenchymal stem cells during long-term culture in vitro Xu Chao, Liao Ji-dong, Liu Jing, Mao Wen-zhe, Li Yang-qiu 2011, 15 (10):  1750-1754.  doi: 10.3969/j.issn.1673-8225.2011.10.009 Abstract ( 348 )   PDF (1287KB) ( 544 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) possess multi-lineage differentiation potential and immunomodulatory effects, and can be made long-term culture in vitro. However, it is still not very clear about the restriction and biological activity feature of MSCs during long-term culture in vitro. OBJECTIVE: To investigate changes of biological activity feature and restriction of human umbilical cord MSCs (hUC-MSCs) for long-term culture in vitro. METHODS: The umbilical cord from cesarean section at term newborns was collected in sterile condition. MSCs were isolated from human umbilical cord by collagenase Ⅱ digestion. The hUC-MSCs were obtained by plastic surface adhesion selection, continuous passage culture by trypsin digestion. The cells surface antigens were identified by flow cytometry, adipogenic and osteogenic ability were determined by specially inducing culture from third generation. A series of different generation cells were collected for following analysis. The cell growth curve was analyzed by MTT assay. The cell cycle and apoptosis rate were measured by flow cytometry. The cell adhesive ability was detected by piece of climbing. RESULTS AND CONCLUSION: The cultured hUC-MSCs phenotype was CD105+/CD29+/CD44+/CD31-/CD34-/CD45-/HLADR-, and could were induced toward osteoblast and adipocyte differentiation under appropriate experimental conditions. There was no significant change in the cellular morphology and the cell growth curve of subculture 3-23 generation. After subculture 28 generation, cell growth became slowly. When subculture 33 generation was compared with subculture 3 generation, the cellular number of G0/G1 phase was decrease 17.9%, and S+G2/M phase and the cell apoptotic rate were increase 103.4% and 316.7%, respectively. Their diversity was statistical significant (P < 0.01). Within subculture 3-18 generation, their diversity of the cellular number of G0/G1 and S+G2/M phase and the cell apoptotic rate were not statistical significant (P > 0.05). Our data show that following increase of subculture frequency, hUC-MSCs proliferate and adherent capabilities of were decrease, while its apoptosis rate increased, in condition of long-term culture in vitro. It’s all biological activity has gradually decreasing trend. The best biological activity period appeared in 3 to 18 generations. Related Articles | Metrics

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Transient expression efficiency of foreign gene in human embryonic stem cells Lu Jun, Chen Zong-feng, Huang Liang-hu, Tan Jian-ming 2011, 15 (10):  1755-1758.  doi: 10.3969/j.issn.1673-8225.2011.10.010 Abstract ( 369 )   PDF (1293KB) ( 422 )   Save BACKGROUND: Technologies designed to allow modification and manipulation of human embryonic stem cells (hESCs) is numerous and various in the complexity of their efficiency, reliability and safety. OBJECTIVE: To compare effect of different enhanced green fluorescent protein (EGFP) expression vectors with different promoter on expression efficiency in human embryonic stem cells (hESCs). METHODS: The H9 cells cultured with non trophoblast were transfected with Fugene HD and lipofectamine 2000, positive colonies were detected under fluorescence microscope, and the expression efficiency were measured using flow cytometry analysis between different transfect vectors such as pCAG-EGFP and pEGFP-N3. RESULTS AND CONCLUSION: Expression efficiencies of pCAG-EGFP and pEGFP-N3 were (42.45±3.32)% and (25.95±1.91)% respectively in Fugene HD transfected hESCs, and they had statistically difference (P < 0.05). It was (1.94±0.18)% and (1.49±0.33)% in lipofectamine 2000. The expression efficiencies of pCAG-EGFP and pEGFP-N3 transfected with Fugene HD were higher than that transfected with lipofectamine 2000 (P < 0.05).These results indicate that Fugene HD have the greater expression efficiency compared to lipofectamine 2000 in hESC line H9; expression efficiency of EGFP driven by CAG promoter is higher than that by CMV promoter. Related Articles | Metrics

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Culture and molecular imaging identification of adipose derived mesenchymal stem cells with stable reporter gene expression in vitro and in vivo Fan Wei-wei, Wang Ya-bin, Zhang Rong-qing, Li Cong-ye, Li Shuang, Cao Feng 2011, 15 (10):  1759-1763.  doi: 10.3969/j.issn.1673-8225.2011.10.011 Abstract ( 253 )   PDF (1693KB) ( 387 )   Save BACKGROUND: Previous studies show that molecular imaging can monitor the stem cell qualitatively and quantitatively at cellular and molecular levels in vivo and hold promise in long-time evaluation of the stem cell. OBJECTIVE: To construct adipose derived mesenchymal stem cells (ADMSCs) with stable reporter genes expression and identify it by reporter gene imaging in vitro and in vivo. METHODS: β-actin-luc transgenic mice were selected firstly.Then ADMSCs were cultured from β-actin-luc mice by modified collagenase method. ADMSCs at passage 3 were identified by flow cytometry. ADMSCs (1×106) were implanted into the muscles of hindlimb of BALB/c nude mice, then tracked and quantified by firefly luciferase (fluc) bioluminescence imaging (BLI) in vitro and in vivo. RESULTS AND CONCLUSION: The transgenic mice stably carried β-actin-luc reporter gene and the ADMSCs were positive for CD90 and CD44, while negative for CD45, CD34 and CD31. ADMSCs consistently expressed the fluc and a robust correlation existed between different cell numbers and fluc average radiance (r2=0.96). After 24-hours engrafted ADMSCs survived and displayed strong BLI signal, which increased rapidly and then decreased smoothly during 0-42 minutes after peritoneal injection of D-luciferin, the peak being (6.92×106±4.11×105) Photons?s-1?cm-2?sr-1 at 21 minutes. Results indicated that ADMSCs of β-actin-luc transgenic mice could highly express the markers of MSCs, stably carry the reporter gene and facilitate the tracking and quantifying by BLI in vitro and in vivo. Related Articles | Metrics

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Influence of monoclonal screening on the purification of primary cultured rat adipose tissue-derived stem cells Liu Jian, Li Li, Ran Jiang-hua, Zhang Sheng-ning, Shao Jian-chun 2011, 15 (10):  1764-1768.  doi: 10.3969/j.issn.1673-8225.2011.10.012 Abstract ( 271 )   PDF (1375KB) ( 314 )   Save BACKGROUND: It was found that a kind of mesenchymal stem cell can be isolated from animal adult adipose tissues, that is, adipose tissue-derived stem cells (ADSCs). However, there were still some problems need to be improved of the traditional isolation methods. OBJECTIVE: To investigate a new method that using monoclonal screening in the purification of primary cultured ADSCs.  METHODS: After isolated the Lewis rat ADSCs with the digestion centrifuge process, the ADSCs were screened with the limited density dilution cloning and condition culture medium screening in the original generation culture. The group with clone screening ADSCs served as the experimental group, without clone screening as the control group. Cellular appearance, growth curve and cell doubling generation time were compared between groups. The surface markers CD49d, CD29, CD44 of ADSCs were determined by flow cytometry. RESULTS AND CONCLUSION: The appearance of ADSCs which underwent clone screening had been homogeneously, with vigorous growing power and short doubling generation time. The growth curve was not changed obviously after many generations. The surface marks CD49d, CD29, and CD44 were expressed in both groups at 3-6 passages. The clone screening purification method is effective, economical purification method of ADSCs. Related Articles | Metrics

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Protein expression of bone marrow mesenchymal stem cells after human vascular endothelial growth factor 165 gene transfection Tian Li, Liang Xiao-peng, Tian Xiao-ye, Yu Xin-chen, Tian Jing 2011, 15 (10):  1769-1772.  doi: 10.3969/j.issn.1673-8225.2011.10.013 Abstract ( 297 )   PDF (679KB) ( 513 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) is the best source of seed cells for tissue engineering, which containing vascular endothelial growth factor 165 (VEGF165) not only has an important significance in angiogenesis and the start of bone repair, but also its sustained release can increase the degree of mineralization of new bone, and enhance the mechanical properties of repair tissue. OBJECTIVE: To observe the hVEGF165 protein function expressed by the gene transfected BMSCs. METHODS: BMSCs from rabbits were isolated and cultured in vitro, and which were purified and identified. Cell surface makers were detected by immunofluorescence assay. And the cultured BMSCs were transfected with the composite of PcDNA3.1-hVEGF165 plasmid and lipofectamine in the ratio of 1/3 mixed liquor, and which were divided into 3 groups: transfection group, empty vector transfection group, and non-transfection group. PcDNA3.1-hVEGF165 plasmid was used to transfect cells in transfection group; pcDNA3.1-empty vector transfection was applied in empty vector transfection group; no treatment was performed in non-transfection group. The expression of exogenous hVEGF165 was detected by ELISA and Western-blot. RESULTS AND CONCLUSION: Compared with empty vector transfection group and non-transfection group, the protein level of VEGF165 in transfection group was significantly higher (P < 0.05), however, there was no significant difference between empty vector transfection group and non-transfection group (P > 0.05). There were significant differences of the protein level of VEGF165 in transfection group (P < 0.05) at different time points. The transfected BMSCs by hVEGF165 can successfully secret the protein of VEGF165. It is indicated that hVEGF165 can be transfected to BMSCs and efficiently express VEGF165 protein of biological activity by using of gene transfection technology. Related Articles | Metrics

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Auxiliary role of origin recognition complex binding site for silencer involved in transcriptional silencing Zhang Xin-min, Wang Hao-tian, Liu Nan, Yu Qun, Yan Wei-qun, Bi Xin 2011, 15 (10):  1773-1776.  doi: 10.3969/j.issn.1673-8225.2011.10.014 Abstract ( 415 )   PDF (506KB) ( 435 )   Save BACKGROUND: Silencer is consisted of origin recognition complex (ORC), RAP1 and other elements. There were a lot of reports on function and relationship between silencers but few researches concerning silencer elements. OBJECTIVE: To study the auxiliary gene silencing effect of ORC to silencer. METHODS: Delete or replace HML-I silencer with ORC binding site with URA3 in the right side or blank sequence without any binding sites to compare expression level of URA3 and analyze the difference of chromatin structure with DNA topological method. RESULTS AND CONCLUSION: When HML-E silencer exited alone, yeast could not grow on FOA plates. But with auxiliary effect of ORC binding site, it was able to grow on the FOA plate, and the proportion of negative super coiled DNA was larger compared with that in the case that HML-E silencer exits alone. Therefore, ORC can interact with HML-E to condense the chromatin structure and enhance gene silencing. Related Articles | Metrics

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The value of morphology and immunological classification and fusion gene chromosome examination in acute leukemia M2 patients Yi Li-ping, Guo Xin-hong 2011, 15 (10):  1777-1782.  doi: 10.3969/j.issn.1673-8225.2011.10.015 Abstract ( 349 )   PDF (566KB) ( 467 )   Save BACKGROUND: Only about 70% of acute leukemia can be classified correctly by traditional morphological classification, simple FAB classification has not met the clinical requirements. OBJECTIVE: To evaluate the diagnostic value and prognostic significance of morphology, immunological classification, fusion gene, chromosome in acute leukemia M2 (AML-M2) patients. METHODS: A total of 76 AML-M2 patients, comprising 40 males and 36 females, who were received treatment, were enrolled from First Affiliated Hospital of Xijiang Medical University. The expression of AML1/ETO fusion gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) technology, karyotype analysis and flow cytometry immunological classification were detected by R-band method, morphology, immunological classification, fusion gene, chromosome were used to examine, continuously observe, and periodically inspect hemogram and other felated inspection. The diagnostic value and prognostical factors were analyzed by the prediction of clear diagnosis or coefficient diagnosis, recurrence with or without the relevant checks. RESULTS AND CONCLUSION: Six patients in 76 cases were not easily classified by morphology. Morphology combined with immunological classification, fusion gene, chromosome can be examined definitely. The myeloid antigen expression has higher remission rate than the stem-ancestor antigen expression. There are across expression by CD19 antigen expression and some acute leukemia antigen expression which has a good prognosing. Seven cases with CD7 antigen expression were completely relieved, and may be more inclined to differentiate into mature cells. The cases which have pure t (8, 21), positive fusion gene have a better prognosis and higher remission rate. The combination of morphology, immunological classification, fusion gene and chromosome can increase diagnosis accuracy and predicting prognosis of AML M2 patients. Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation combined with Piracetam for repair of spinal cord injury in rats  Jia Hon-wei, Li Yan-ying 2011, 15 (10):  1783-1788.  doi: 10.3969/j.issn.1673-8225.2011.10.016 Abstract ( 334 )   PDF (741KB) ( 467 )   Save BACKGROUND: Simple stem cell transplantation for the repair of spinal cord injury (SCI) is not ideal, mainly because edema, ischemia, hypoxia of nerve tissue in damage area after SCI injury caused secondary injury. OBJECTIVE: To investigate the effect of bone mesenchymal stem cell (BMSC) transplantation and Piracetam on repair of rat’s spinal cord injury (SCI). METHODS: SCI models of female Wistar rats were prepared with reference to Allen punch method, and which were randomly divided into 3 groups: SCI group, BMSC transplantation group, BMSC transplantation + Piracetam group. At 1, 2, 4, 6, 8 weeks post-injury, all animals were evaluated of the hind limb behavior with BBB score and inclined plane test. Four weeks post-transplantation, pathological sections hematoxylin-eosin staining was performed. SRY-PCR was used to detect the SRY gene which is a specific marker of Y chromosome, so that whether transplanted BMSC is survival. Eight weeks post-transplantation, horseradish peroxidase (HRP) tracing observation was performed, and the regeneration of axonal was observed according to transmission electron microscope. RESULTS AND CONCLUSION: At 4 weeks after injury, the motor functions of hind limbs of rats between BMSC transplantation group and BMSC transplantation+ Piracetam group had significant recovery, the outcome of BMSC transplantation+ Piracetam group surpassed the BMSC transplantation group’s (P < 0.05). The SCI group’s outcome was similar to BMSC transplantation group and BMSC transplantation + Piracetam group, but its degree was lighter than the others’. Pathological simple SCI group has not seen the neural neuraxial through, a small amount of nerve axon-like structure could be seen in BMSC transplantation group, more nerve axon-like structure could be seen in BMSC transplantation+ Piracetam group.The SRY expression was detected in BMSC transplantation group and BMSC transplantation + Piracetam group, while SRY gene was not detected in SCI group. HRP-labeled neurofibra was found in spinal cord of BMSC transplantation+ Piracetam group, some in BMSC transplantation group but little in SCI group, there was significant difference (P < 0.05). Neonatal unmyelinated nerve fiber and myelinated nerve fiber could be seen in central transverse plane in BMSC transplantation group and BMSC transplantation + Piracetam group under transmission electron microscope. The effect of structural and functional recovery after SCI in BMSC transplantation + Piracetam group was significantly superior to BMSC transplantation group. BMSC transplantation combined with Piracetam has synergistic effects. Related Articles | Metrics

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Treatment of hypoxic-ischemic brain damage by hyperbaric oxygen combined with bone marrow mesenchymal stem cells transplantation   Zhang Ting-yong 2011, 15 (10):  1789-1793.  doi: 10.3969/j.issn.1673-8225.2011.10.017 Abstract ( 267 )   PDF (735KB) ( 400 )   Save BACKGROUND: The repair effect of single bone marrow mesenchymal stem cells (BMSCs) transplantation on treating cerebral infracted tissues are poor, which need to be combined with drugs and bioengineered materials. OBJECTIVE: To determine whether the hyperbaric oxygen (HBO) treatment can enhance curative effects of BMSCs transplantation on hypoxic-ischemic brain damage in rats. METHODS: Rat BMSCs were cultured by the suspension culture in vitro. The models of cerebral infarction were made by middle cerebral artery occlusion (MCAQ). Rats were divided randomly into control group, BMSCs group and HBO+BMSCs group. The neurobehavioral scores were determined after 24 hours, 3 days and 1, 2 weeks after MCAQ. After 2 weeks, RT-PCR was used to assess the GAP-43 gene expression. The recovery condition was detected by BrdU immunohistochemistry and pathology. RESULTS AND CONCLUSION: The neurobehavioral scores were significantly lower in the HBO+BMSCs group than those in control group and BMSCs group at 1 week after MCAQ (P < 0.05). The expression of GAP-43 mRNA was highest in the HBO+BMSCs group, a few in the BMSCs group and lowest in the control group (P < 0.05). The number of neurons in the HBO+BMSCs group was higher than that in BMSCs group and control group (P < 0.05). Hyperbaric oxygen treatment combined with BMSCs transplanted by vein into the hypoxic-Ischemic brain damage tissues can significantly improve the neurological function in the rats with MCAQ, the curative effects is superior to single BMSCs transplantation. Related Articles | Metrics

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Effects of long term intranasal delivering umbilical cord blood mononuclear cells on cerebral infarct volume and behaviors in rats Zeng Xian-zhi, Shen Hui-juan, Wang Yang, Zhu Qun-e, Guo Lian-jun, Ye Xian-cai 2011, 15 (10):  1794-1798.  doi: 10.3969/j.issn.1673-8225.2011.10.018 Abstract ( 233 )   PDF (1662KB) ( 321 )   Save BACKGROUND: It was a new non-invasive method for intranasal delivering cell or cytokine into the brain, which could overcome the peripheral side-effect of other delivering methods. OBJECTIVE: To explore the effect of long term intranasal delivering human umbilical cord blood mononuclear cells (UCBMCs) on changes in behaviors and infarct volume of brain infarct in rats. METHODS: Middle cerebral artery of Wistar rat was directly cut to establish models of cerebral infarction. Rat models with equal neurological score were randomly assigned to control and mononuclear cell groups. In the mononuclear cell group, UCBMCs of the third generation were intranasally injected. In the control group, phosphate buffer saline was infused, once a day, totally 6 weeks. Behavior was assessed once a week. Brain infarct volumes were quantified following the last behavior assessment. RESULTS AND CONCLUSION: Compared with control group, modified neurological severity score was improved in the mononuclear cell group at 3 weeks after treatment, and lasted at 5 weeks (P < 0.05, P < 0.01). In the mononuclear cell group, the mean latency in Morris Water Maze Test shortened significantly at 1-5 weeks following treatment (P < 0.05, P < 0.01), and the effect was obvious over time. Identical to behavior improvement, the cerebral infarct volume in the mononuclear cell group (9.15±4.36)% was smaller compared with (30.56±4.65)% (P < 0.01). Early and long term of intranasal delivering UCBMCs to treat brain infarct model rats could improve their behavior score and shrink their infarct volume, which could be considered as a simple and feasible method to treat brain infarct. Related Articles | Metrics

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Allogeneic bone marrow mesenchymal stem cells transplantation in treatment of acute liver failure He Jin-qiu, Yang Ling-ling, Lei Yan-chang, Yang Wen-long, Zhou Xi-jing 2011, 15 (10):  1799-1802.  doi: 10.3969/j.issn.1673-8225.2011.10.019 Abstract ( 267 )   PDF (1347KB) ( 409 )   Save BACKGROUND: Effective therapies are deficient in treating acute liver failure (ALF). At present, there is no effective method. Bone mesenchymal stem cells (BMSCs) can differentiate into hepatocyte-like cells and participate in liver regeneration and repair, which provides a new manner for treatment of ALF. OBJECTIVE: To evaluate the effect of BMSCs transplantation on the ALF in rats, to provide empirical study foundation for future clinical application. METHODS: The BMSCs were cultured and purified by the whole marrow-cultured method. A total of 30 healthy Sprague-Dawley rats were randomly assigned to three groups. Normal control group: no treatment. ALF group and transplantation group: ALF models of rats were developed by intraperitoneal injection with CCL4. 24 hours following model induction, physiological saline and an equal volume of BMSCs were injected via the caudal vein. Finally, liver function was tested at 1, 2, 3, 7 days intervals and histopathologic examinations were performed. RESULTS AND CONCLUSION: Following BMSCs transplantation, the survival rates of the rats were increased to 70%, compared to the ALF group (20%), and the difference was statistically significant (P < 0.05). The activities of alanine aminotransferase and aspartate aminotransferase were significantly lower in the transplantation group compared with ALF group (P < 0.05). In pathohistology, the degeneration and necrosis of hepatic cells and degree of inflammation were less obvious than that of ALF group. Hence, BMSCs transplantation via caudal vein could elevate the survival rate of ALF rats, improve hepatic function and alleviate the degree of cell necrosis, and could help to treat ALF in rats. Related Articles | Metrics

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Reconstruction of hepatic functions after bone marrow mesenchymal stem cells transplantation Liu Fang, Chen Shao-hong, Liu Qing-bo, Fan Xin-juan, Tang Fang, Zhao Guo-qiang 2011, 15 (10):  1803-1808.  doi: 10.3969/j.issn.1673-8225.2011.10.020 Abstract ( 229 )   PDF (1839KB) ( 456 )   Save BACKGROUND: Cell transplantation is an effective method for the treatment of hepatic function failure. However, ideal cell source and the best way to transplant has been the target of the ongoing research in laboratories. OBJECTIVE: To observe the effect and optimal transplantation way of bone marrow mesenchymal stem cells (BMSCs) transplantation on hepatic function reconstruction with acute hepatic failure in rats. METHODS: D-galactosamine (D-gal) was served as hepatic toxic agent, female SD rats with acute hepatic failure model was constructed. Male rats BMSCs as cell source was implanted hepatic failure rats through groin vein and intrasplenic transplantation. Many hepatic function blood biochemical indices were detected dynamically. Sry gene was served as molecule marker, the survival status of BMSCs in hepatic was detected by PCR technique. RESULTS AND CONCLUSION: BMSCs through groin vein and intrasplenic transplantation can lower the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBIL), improve the ability of albumin (ALB) synthesis, ameliorate hepatic function, and reduce the death rate of acute hepatic failure rats. The death rate in the group of groin venous transplantation was lower than that in the intraspleen transplantation group. Sry gene positive cell was detected in hepatic of receptor rats at 7-30 days after transplantation; however, sry gene positive cell was not detected in hepatic of receptor rats in corresponding control group. The results showed that BMSCs can ameliorate hepatic function of acute hepatic failure rats. BMSCs can survive in the hepatic of receptor rats. The operation of groin venous transplantation is simple and convenient; the death rate is lower. So groin venous transplantation is a better way to treat acute hepatic failure rats. Related Articles | Metrics

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Neural stem cells transplantation for the treatment of spinal cord injury Liu Ning, Hu Dian-lei, Xu Tao, Yu Sheng-hui, Sheng Wei-bin 2011, 15 (10):  1809-1813.  doi: 10.3969/j.issn.1673-8225.2011.10.021 Abstract ( 295 )   PDF (1704KB) ( 462 )   Save BACKGROUND: Many studies showed that neural stem cells (NSC) transplantation can promote functional improvements in spinal cord injury (SCI) rats. However, the activity of proliferation, differentiation and migration of the transplanted NSCs in the injured zone remain poorly understood. OBJECTIVE: To investigate the effects of NSCs transplantation on the function recovery of rat hind limb with SCI. METHODS: SD rats were randomized into 3 groups after the animal models of spinal cord completely transsected at T10 had been established for 1 week. Control injuried group only exposed spinal cord; 10 μL DMEM/F12 fluid and 10 μL NSCs (1.0×109/L) were transplanted into the caulal zone of the spinal cord in the transplantation control and NSCs groups, respectively. The function repair was evaluated by Bundle branch block (BBB) score and pathology; Imunnoflurore-scence technique was used to detect the surviveal, migration and differentiation of the Brdu-labeled cells in vivo. RESULTS AND CONCLUSION: A cultivation of NSCs derived from rat hampocampus on proliferation and differentiation in vitro was established successfully; The BBB scores of the transplantation control and NSCs groups were increased with time prolonged, which was more greater in the NSCs group from 2 weeks after transplantation (P < 0.05); NSCs could continue survive and migrate into the injured parts in vivo after transplantation, and some of them could differentiated into NF-200 and the glial fibriuary acidic protein positive cells that had characteristics of neuron or astroglia. It suggested that NSCs transplantation is an effective method to repair SCI in rats. Related Articles | Metrics

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Expressions of nuclear factor κBp65 and proliferating cell nuclear antigen in injured carotid artery balloon after bone marrow mesenchymal stem cells transplantation Liu Zhi-jiang, Shi Bei, Xu Guan-xue, Zhao Ran-zun, Shen Chang-yin, Wang Dong-mei, Wang Zheng-long 2011, 15 (10):  1814-1818.  doi: 10.3969/j.issn.1673-8225.2011.10.022 Abstract ( 281 )   PDF (1701KB) ( 432 )   Save BACKGROUND: Inflammatory reaction plays an important role in the incidence of restenosis after percutaneous transluminal coronary intervention. Recent studies found that bone marrow mesenchymal stem cells (BMSCs) have strongly characteristics of immunological regulation. However, BMSCs on the inflammatory reaction in vascular injury is rarely reported. OBJECTIVE: To investigate effects of BMSCs transplantation on expression of nuclear factor κBp65 (NF-κBp65) and proliferating cell nuclear antigen (PCNA) after carotid artery balloon injury of rabbits, and to analyze its correlation with intimal proliferation. METHODS: Thirty-six rabbits were prepared for carotid artery atherosclerotic stenosis models and were randomly divided into the control group and BMSCs transplantation group. BMSCs pre-labled by DAPI was infused into rabbits by the ear vein in the BMSCs transplantation group, and the same amount of PBS solution was infused in the control group. The plasma tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were detected by ELIAS in prior to model preparation and at 7, 14, 28 days after transplantation. The NF-κBp65 and PCNA expressions in vessels tissues were detected by immunohistochemical analysis at 14 days after transplantation, and vascular morphological changes were determined by hematoxylin-eosin staining at 28 days after transplantation. RESULTS AND CONCLUSION: The expression of NF-κBp65 and PCNA in the BMSCs transplantation group decreased significantly compared with control group at 14 days after transplantation (P < 0.05). At 7, 14, and 28 days after transplantation, the plasma TNF-α and IL-6 levels in the BMSCs transplantation group were significantly lower than those in the control group (P  < 0.05). The intimal area, the ratio of the intima/media area and the luminal stenosis ratio were significantly decreased in the BMSCs transplantation group than control group at 28 days after transplantation (P  < 0.05). BMSCs is capable to decrease the inflammatory reaction of injured vessels and down-regulate PCNA expression by inhibiting NF-κBp65 in injured vessels, and BMSCs may be good for inhibition of injured vessels intimal proliferation. Related Articles | Metrics

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A method of pancreatic stem cells culture and amplification in vitro  Kou Ya-l1, Hu Li-xia, Li Qin, Zhao Wen-jing, Chen Wei-ping 2011, 15 (10):  1819-1822.  doi: 10.3969/j.issn.1673-8225.2011.10.023 Abstract ( 327 )   PDF (1152KB) ( 381 )   Save BACKGROUND: Pancreatic stem cells are differentiated easily when cultured in vitro, so it is difficult to obtain a sufficient number of pancreatic stem cells. OBJECTIVE: To explore a simple method which pancreatic stem cells were amplified and cultured in vitro. METHODS: The pancreases of newborn Kunming mice were isolated. After culturing in vitro for 3-4 days, the colony growing pancreatic stem cells were emerged in fibroblasts. The number of fibroblasts was reduced by 0.02% EDTA; the fibroblasts were cultured continuously after their proportions were reduced. When the cells of confluent culture reached 80% of bottom, repeat 3 or 4 times, the number of pancreatic stem cells was gradually increased. The specific marker (Nestin) of pancreatic stem cells was detected. Differentiation of pancreatic stem cells was induced by nicotinamide. Islet-like cell clusters were detected by dithizone (DTZ) staining. RESULTS AND CONCLUSION: After the number and proportion of fibroblasts were reduced, the pancreatic stem cells presented active proliferative ability. The number of the pancreatic stem cells was significantly increased. The amplificated pancreatic stem cells maintained their properties, including round appearance with single nucleus, strong diffraction, and expression (Nestin). After induced by nicotinamide, the amplificated pancreatic stem cells were differentiated to islet-like cell clusters with positive DTZ staining. The results demonstrated that the proportion and number of fibroblasts were reduced which were beneficial for proliferation of pancreatic stem cells. A large number of pancreatic stem cells were obtained. The amplificated pancreatic stem cells with undifferentition condition still had the potential of differentiation to pancreatic islet under suitable induction condition. Related Articles | Metrics

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Biological characteristics of human chorionic-derived mesenchymal stem cells Zhang Rui-ting, Han Zhi-bo, Wang Tao, Meng Lei, Liu Zhi-peng, Li Kui, Ji Yue-ru, Yang Zhou-xin, Li Yang-qiu, Han Zhong-chao 2011, 15 (10):  1823-1826.  doi: 10.3969/j.issn.1673-8225.2011.10.024 Abstract ( 399 )   PDF (1249KB) ( 638 )   Save BACKGROUND: Bone marrow is major source of mesenchymal stem cells (MSCs), but the quantity and quality of MSCs in bone marrow have been shown to be gradually reduced with the increase of age. Therefore, it’s very important to find a new source of stem cells. OBJECTIVE: To study a new source of MSCs isolated from human chorionic villous and to study its biological characteristics. METHODS: Umbilical cord and amniotic membrane were isolated from clean placenta; the remaining placenta tissue underwent primary culture and subculture. Tandem repetitive sequence (TRS) analysis was used to detect whether the MSCs obtained from placenta is chorionic villous original. Cell growth pattern was detected by MTT assay. Flow cytometry was applied to analyze cell phenotype and stem cell markers. The different differential medium was used to detect its multi-directional differentiation ability. The immunomodulatory properties of these chorionic-derived cells were evaluated by coculturing the cells with peripheral blood mononuclear cells stimulated by phytohemagglutinin (PHA), and the level of interferon-γ (IFN-γ) was detected by enzyme-labeled immunosorbent assay (ELISA). RESULTS AND CONCLUSION: TRS analysis shows that these chorionic-derived cells are derived from human chorionic. They are adherent cells with typical fibroblast morphology. Flow cytometric analysis reveals that human chorionic-derived cells are consistently positive for MSCs labeled with CD73, CD29, CD44, CD90 and CD105, and negative for the hematopoietic markers CD45, CD11b and CD34. The cells can differentiate into osteoblasts and adipogenesis under different conditioned medium cultured conditions. These results proved that these cells are human chorionic-derived MSCs. The human chorionic-derived MSCs can inhibit peripheral blood mononuclear cell stimulated by PHA to secrete γ-interferon. Related Articles | Metrics

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Human embryonic stem cells maintained and cultured in serum-free medium mTeSR?1 and induced into endothelial cells differentiation Li Xiang-dong, Wang Ji-wen, Wei Guo-feng 2011, 15 (10):  1827-1831.  doi: 10.3969/j.issn.1673-8225.2011.10.025 Abstract ( 325 )   PDF (1562KB) ( 472 )   Save BACKGROUND: Containing fetal bovine serum medium is used in the traditional culture and amplification method of human embryonic stem cells (hESCs), and this method relys on feeder layer cell culture and significantly limits culture formula of stem cells in vitro. In addition, the intervention of heterologous serum components significantly increases pathogen contamination and the probability of immune rejection. OBJECTIVE: To elucidate the feasibility of serum-free medium mTeSR?1 on the hESCs long term cultured in vitro and to establish a platform of induced hESC differentiate into endothelial cells. METHODS: Serum-free medium mTeSR?1 was applied to culture in vitro and amplificate hESCs line H9 by non- feeder layer cell dependent. After more than 40 times passage in vitro, the growth morphology of hESCs was observed by inverted microscope, and their phenotype was evaluated by immunofluorescence staining method. Moreover, a conditioned medium was utilized to induce hESCs line H9 to differentiate into endothelial cells. The phenotype and function of ESC-derived endothelial cells were assayed by immunofluorescence staining, quantitative RT-PCR, and low-density lipoprotein (LDL) uptaking experiment. RESULTS AND CONCLUSION: mTeSR?1 medium can support hESCs line H9 long term amplification in vitro by non- feeder layer cell dependent and maintain its potential of undifferentiated stem cells. When the hESCs were cultivated under a conditioned medium with vascular endothelial cell supplementation, the cells were induced differentiation into H9 endothelial-like cells. These cells not only one of important surface markers of endothelia cells (kdr, pecam) and expressed CD31, but also uptake LDL, formed vascular-like structure during the differentiation. The system of culture and induced differentiation experiment provided can support the proliferation and differentiation behavior of ESCs. Related Articles | Metrics

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Effects of fasudil and RhoA silencing on neural stem cell proliferation Wang Bing-qian, Wang Dong, Zhang Jian-jun, Wang Yong-xin, Zhu Guo-hua, Du Guo-jia, Dang Mu-ren 2011, 15 (10):  1832-1836.  doi: 10.3969/j.issn.1673-8225.2011.10.026 Abstract ( 302 )   PDF (1278KB) ( 344 )   Save BACKGROUND: Rho and its relevant molecules play important roles in growth, differentiation, extension and synapse formation of nerve axons. To block and inhibit RhoA/ROCK pathway can promote the proliferation and growth of neural stem cells. OBJECTIVE: To observe the effects of Rho kinase inhibitor fasudil and RNAi-mediated RhoA gene silencing on the proliferation of neural stem cells in rats. METHODS: Neural stem cells of Wistar fetal rats were cultured in vitro, and were divided into six groups: blank control group, 5, 10, 15, 20 μmol/L fasudil groups, and siRNA silencing RhoA gene group. On day 3 following intervention, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the expression of RhoA mRNA and RhoA protein of neural stem cells in each group. Cellular proliferation was determined by MTT assay. The cell cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION: RhoA gene and protein expression was significantly lower in the 15, 20 μmol/L fasudil groups and siRNA silencing RhoA gene group compared with 5, 10 μmol/L fasudil groups and blank control group (P < 0.05). Cell growth speed was significantly faster compared with the 5, 10 μmol/L fasudil groups and blank control group (P< 0.05); cell number was reduced in the cell cycle G0/G1 (P < 0.05) and increased in S phase (P < 0.05). At 20 μmol/L fasudil, cell effect was not enhanced with the increased concentration. No significant difference was determined compared with 15 μmol/L fasudil group (P > 0.05). No significant difference was detected between 15, 20 μmol/L fasudil groups and siRNA silencing RhoA gene group (P > 0.05). Results suggest that Rho kinase inhibitor fasudil and RNAi-mediated RhoA gene silencing can promote the proliferation of neural stem cells in vitro. The optimal concentration of fasudil was 15 μmol/L. Related Articles | Metrics

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Optimization of the isolation and cultivation method of human amniotic fluid-derived mesenchymal stem cells Guo Su-hong, Yuan Chun-hua, Hou Yi-jü, Liu Hui-ling 2011, 15 (10):  1837-1840.  doi: 10.3969/j.issn.1673-8225.2011.10.027 Abstract ( 305 )   PDF (1272KB) ( 321 )   Save BACKGROUND: There are different methods of in vitro isolating culture for human amniotic fluid-derived mesenchymal stem cells (AF-MSCs). To gain most AF-MSCs is still difficult. OBJECTIVE: We had isolated 80 cases AF, and compared different in vitro culture methods for optimization and bolted a better method to establish foundation of AF-MSCs in clinical widespread application. METHODS: Amniotic fluid was collected from full-term deliveries and artificial labors under aseptic condition. The amniotic fluid cells were isolated by centrifugation. The influences of different pregnancy ages, culture media, incubation densities and the time points of the first medium change on the growth of AF-MSCs were analyzed. The cell metabolism was determined and biological characteristics of AF-MSCs were observed by cytochemical staining. RESULTS AND CONCLUSION: When the other conditions were the same, AmnioMAXⅡcomplete medium was the best medium. 5×104/cm2 was the best planting density; the best time for the first medium change was on the sixth day. The achievement ratio of amniotic fluid from fetation of intermediate stage (14-20 weeks) was higher than that of later fetation (21-36 weeks) and full-term deliveries (37-42 weeks) when the other conditions were the same. AF-MSCs highly expressed periodic acid-schiff, acid phosphatase, α-naphthol acetate esterase (α-NAE) and weakly expressed neutrophil alkaline phosphatase, but no peroxidase and Sudan black was found. Results suggest that we have found the optimized method of culturing AF-MSCs. Related Articles | Metrics

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CD28 mRNA expression in peripheral blood mononuclear cells of patients with acute myeloid leukemia Wang Ya-nan, Shi Jin-fang, Gu Guo-hao 2011, 15 (10):  1841-1844.  doi: 10.3969/j.issn.1673-8225.2011.10.028 Abstract ( 307 )   PDF (1007KB) ( 314 )   Save BACKGROUND: Recent studies have found that there is difference in costimulatory molecule CD28 protein expression on the surface of peripheral blood T cells in tumor patients, which suggested that abnormal costimulation pathway may be associated with the occurrence and progress of malignant tumor. OBJECTIVE: To investigate the costimulatory molecule CD28 mRNA expression in peripheral blood mononuclear cells of acute myeloid leukemia patients. METHODS: A total of 80 patients with acute myeloid leukenmia patients were selected, including 7 cases of M0 type, 6 cases of M1 type, 18 cases of M2 type, 15 cases of M3 type, 17 cases of M4 type, 9 cases of M5 type, and 8 cases of M6 type. According to acute leukemia curative effect criteria, 80 patients were assigned to complete healing group, remission group and non-remission group. Using Taqman probe, real-time fluorescent quantitation polymerase chain reaction was used to detect CD28 mRNA expression in peripheral blood mononuclear cells of 80 patients and 76 healthy persons. RESULTS AND CONCLUSION: CD28 mRNA expression was lower in the M1, M3, M4 subtype in peripheral blood mononuclear cells of patients with acute myelocytic leukemia compared with healthy population (P < 0.05). CD28 mRNA expression in non-remission group was lower than healthy population (P < 0.05). No significant difference in CD28 mRNA expression was detected between the complete healing and remission groups and healthy population. Results suggested that acute myeloid leukemia patients possess defects in peripheral blood mononuclear cells CD28 mRNA expression which associates with the clinical stage, disease progress and prognosis. Related Articles | Metrics

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Chinese herbs for inducing osteogenic differentiation of bone marrow mesenchymal stem cells  Yang Feng, Tang De-zhi, Bian Qin, Wang Yong-jun, Shi Qi 2011, 15 (10):  1847-1850.  doi: 10.3969/j.issn.1673-8225.2011.10.030 Abstract ( 332 )   PDF (753KB) ( 521 )   Save BACKGROUND: Some Chinese herbs can induce osteogenic differentiation of mesenchymal stem cells (MSCs). The potency of MSCs differentiate into osteogenesis has a theoretical communication with traditional Chinese medicine (TCM) treatment of osteoporosis, fractures, bone necrosis, bone defects and other bone-related diseases. OBJECTIVE: To understand the researching status of osteogenic differentiation of MSCs induced by Chinese herbs so as to establish further researching foundation. METHODS: CNKI database (2000-01/2010-06) and PubMed (2000-01/2010-06) were retrieved by the first author, with the key words of “Chinese herb, mesenchymal stem cells, osteogenic differentiation” in Chinese and in English. The literatures were limited to Chinese and English languages. Literatures concerning TCM monomer, single Chinese herb, Chinese herbal compound, and serum containing Chinese herb inducing osteogenic differentiation of MSCs in vivo and in vitro, human and animal were included. Repetitive studies were excluded. RESULTS AND CONCLUSION: A total of 32 literatures were included, including 2 documents addressing research background of MSCs, 5 single Chinese herbs (4 kidney-nourished herbs and 1 Qi-nourished herb), 10 Chinese herbal compound (6 kidney-nourished formulas and 4 kidney-nourished and activating blood formulas), 15 effective components of Chinese medicine. The kidney-nourished Chinese herbs, Qi- nourished herbs and activating blood herbs can induce MSCs into osteogenic differentiation, but the kidney-nourished Chinese herbs are majority. It explores the scientific value of the theory of “kidney control bone” and supply more resources of seed cells for a large number of amplification of MSCs in vitro, osteogenic differentiation and bone tissue engineering. Related Articles | Metrics

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Action and mechanism of umbilical cord blood stem cells transplantation for treatment of myocardial infarction  Du Dan, Zhang Ming 2011, 15 (10):  1851-1854.  doi: 10.3969/j.issn.1673-8225.2011.10.031 Abstract ( 310 )   PDF (453KB) ( 434 )   Save BACKGROUND: Many basic and clinical studies have demonstrated that stem cell transplantation can improve cardiac function in patients with heart failure following myocardial infarction. OBJECTIVE: To summarize the action and mechanism of umbilical cord blood stem cells (UCBSCs) transplantation for treatment of myocardial infarction. METHODS: The first author retrieved PubMed Database, CNKI and Wanfang Database for articles of basic research on UCBSCs transplantation for treating myocardial infarction published from 2000 to 2009. RESULTS AND CONCLUSION: Stem cell transplantation for treating acute myocardial infarction exhibits advantages compared with traditional therapeutic method. A series of present basic researches have confirmed that human UCBSCs can be an ideal cell source, but these studies are in exploratory study stage and many problems should be solved, such as in vitro isolation and culture of human UCBSCs, optimal transplant therapeutic time, favorable transplant pathway, optimal transplant number, survival rate of transplanted cells, differentiation into myocardial cells or not posttransplantation, differentiation degree, therapeutic effects and safety. Related Articles | Metrics

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Research and application of induced pluripotent stem cells in nervous system diseases Li Jia-su, Yao Zhong-xiang 2011, 15 (10):  1855-1858.  doi: 10.3969/j.issn.1673-8225.2011.10.032 Abstract ( 320 )   PDF (602KB) ( 477 )   Save BACKGROUND: Induced pluripotent stem cells (iPSCs) can differentiate into many kinds of expected neural cells to treat nervous system diseases which lack of neural cells, and play a relevant functional role. Recently, lots of encouraging progresses have been made. How to improve the efficiency and ensure graft substitute security, and be better applied to the clinical research has become a hot spot. OBJECTIVE: To review the recent research and application of iPSCs in Parkinson’s disease, spinal cord injury, motor neuron disease and other nervous system diseases. METHODS: We searched PubMed database (2006-01/2010-09) using “induced pluripotent stem cells, neurological diseases” as research words, and searched the Chinese Biomedical Literature Database (2006-01/2010-09) using “induced pluripotent stem cells” as the key words. RESULTS AND CONCLUSION: A total of 149 articles on induced pluripotent stem cells were collected, including 17 Chinese articles, and 132 English articles. By reading the title and summary, articles that have nothing to do with this article and those with repetitive content were excluded. Finally, a total of 25 articles were reviewed. The iPSCs as a potential new treatment can be used as donor cells and diseases of the carrier, and have greatly potential value in Parkinson’s disease, spinal cord injury, muscular atrophy motor neurons and other nervous system diseases. Basic and clinical researches have made great achievements, but induction efficiency and transplantation safety are still major obstacles to its application and development. Related Articles | Metrics

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Neural stem cells and exercise-induced neurological disease Zeng Yu 2011, 15 (10):  1859-1862.  doi: 10.3969/j.issn.1673-8225.2011.10.033 Abstract ( 393 )   PDF (613KB) ( 303 )   Save BACKGROUND: Neural stem cells are characterized by multi-directional differentiation potential, self-renewal, migration, low immunity, which has been widely used in clinical practice. However, there are rare researches addressing neural stem cells in the field of sports medicine for sports injury prevention. OBJECTIVE: To explore the clinical applications of neural stem cells through its biological characteristics, and provides a theoretical basis for sports injury prevention and treatment. METHODS: The articles related to neural stem cells and exercise-induced neurological disease in CNKI database and PubMed database from January 1997 to October 2010 were retrieved with the key words of “Neural Stem Cell, Sports Medicine, Denervation Muscle Atrophy, Peripheral Nerve Injury” in Chinese or in English. The contents of articles concerning neural stem cells and exercise-induced neurological disease, and recently published or published in the authoritative magazines in the same field were selected. A total of 262 literatures were obtained from computer screen, and 31 documents of them were involved for summarization according to inclusion criteria. RESULTS AND CONCLUSION: Neural stem cells are characterized by multi-directional differentiation potential, self-remaining and renewal, low immunity, migration, and wide variety of sources, which provide a more broad application prospects for the treatment of neurodegenerative diseases, exercise-induced skeletal muscle atrophy, and promote the sport of the regeneration of peripheral nerve injury. But because of the limited basic research, its mechanism of action, induced differentiation, migration need to be confirmed. Related Articles | Metrics

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Effect and mechanism of bone marrow mesenchymal stem cells on the treatment of early avascular necrosis of the femoral head Wang Kun, Gong Yue-kun, Li Biao, Ao Xiao-jun 2011, 15 (10):  1863-1866.  doi: 10.3969/j.issn.1673-8225.2011.10.034 Abstract ( 307 )   PDF (675KB) ( 476 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have great self-proliferative capacity and pluripotent differentiation, so it becomes a hot spot of the treatment of early avascular necrosis of the femoral head (ANFH). OBJECTIVE: To summarize the bionomics of BMSCs and its effect and mechanism on the treatment of early ANFH. METHODS: The literatures were related to the bionomics of BMSCs and its effect and mechanism on the treatment of early ANFH from PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) between January 1992 and October 2010, CNKI database (http://www.cnki.net/index.htm) between January 2000 and October 2010 were retrieved. The index words were “femur head necrosis, mesenchymal stem cell, cell culture, therapy, overview” both in English and in Chinese. Outdated and repetitive researches were excluded, and 30 literatures were included for summarization. RESULTS AND CONCLUSION: Using osteogenic action and angiogenesis directly treated lesion location of ANFH, so as to achieve the purpose of the treatment of early ANFH, which is the hot spot of the treatment of early ANFH currently. In future, we should further analyze BMSCs in the potential and influential factor of proliferation and differentiation of ischemic ANFH, the prevention of restenosis, the time of bone defects repair and survival and differentiation factors of stem cells after BMSCs transplantation, such as local infection and eneral immunity, and whether stem cells can flow to other sites with the impact of the blood flow during interventional therapy, these problems are still to be further solved. Related Articles | Metrics

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Stem cells involved in the tissue repair of sports injury Yang Bo, Liang Jing-qun, Liu Tao, Wang Wang, Zhang Xiang 2011, 15 (10):  1867-1870.  doi: 10.3969/j.issn.1673-8225.2011.10.035 Abstract ( 318 )   PDF (663KB) ( 366 )   Save BACKGROUND: Previous studies demonstrated that mesenchymal stem cells (MSCs) have the potential of multi-directional differentiation, and applied to many field. Stem cells therapy will be a revolutionary progress for tissue injury repairing. OBJECTIVE: To summarize the situation and new progress of soft tissue injury with stem cells therapy at home and abroad. METHODS: The articles related to stem cells involved in the treatment of soft tissue injury in CNKI database and Elsevier database from January 2000 to September 2010 were retrieved by computer with the key words of “stem cells, treatment, soft tissues injury” in Chinese and in English. The content of articles related to stem cells therapy, and recently published or published in authoritative magazines in the same field were selected, and 36 documents of them were involved for summarization. RESULTS AND CONCLUSION: The competitive state of athletes was seriously affected by tendon, skeletal injury in sports, and it is difficult to radical cure, often recurrent attacks. The research of stem cells technology has a great significance in the treatment of injury of athletes. At present, MSCs as a basis of cells therapy is becoming a hot spot and most forward in the field of regenerative medicine research. Therefore, MSCs as the most important source of adult stem cells has a vital application prospect. Related Articles | Metrics

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Myofibroblasts and activated fibroblasts in tumor stroma Yi Ming, Huang Rong, Zhang Jin-feng, Li Shu-zhong 2011, 15 (10):  1871-1874.  doi: 10.3969/j.issn.1673-8225.2011.10.036 Abstract ( 296 )   PDF (572KB) ( 604 )   Save BACKGROUND: Myofibroblasts has always been a controversial topic since was found in 1971. Recent studies suggest that the cells may play an important role in cancer occurrence and development process. OBJECTIVE: To explore the biological characteristics of myofibroblasts and the role of tumor stromal fibroblasts in tumor growth and invasion. METHODS: Literatures about myofibroblast were retrieved from Wanfang database and Pubmed database (1990-01/2010-01) by the first author. The index words were “myofibroblast; tumor; TGF-β; aggressiveness” both in English and Chinese, respectively. The articles related to myofibroblasts and tumor stromal fibroblasts were selected, articles in the same field published in recent years or in authorized journals. A total of 116 literatures were primarily selected, and 41 documents of them were involved for summarization according to the inclusion criteria. RESULTS AND CONCLUSION: Myofibroblasts have the characteristics of smooth muscle cells and fibroblast, tumor interstitial myofibroblasts is the regulation of cell-derived cytokines from fibroblasts transformed by directly stimulating the proliferation and survival of cancer cells to promote tumor growth, and by hydrolysis cellular matrix and stimulates tumor cell motility to occur invasion. The emergence of myofibroblasts contributes to the early diagnosis of tumors and to explore new ways of cancer treatment. Related Articles | Metrics

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miRNA and biological behavior of stem cells Zhang Zi-qiang, Liu Yu-mei, Deng Wen, Lü Qiong-xia 2011, 15 (10):  1875-1878.  doi: 10.3969/j.issn.1673-8225.2011.10.037 Abstract ( 242 )   PDF (576KB) ( 475 )   Save BACKGROUND: In the past, the research about stem cells was focused on the genes. But now, more and more evidence indicate that miRNA plays an important regulatory role in the self-renewal and differentiation process of stem cells. OBJECTIVE: To introduce the formation of miRNA and its effect on the self-renewal and multi-directional differentiation of stem cells. METHODS: A computer based online search of Elsevier database (2000-01/2010-05) was performed with the key words “microRNA, stem cell” in English. Simultaneously, “microRNA, stem cell” in Chinese was used to search CNKI database (2000-01/2010-05). RESULTS AND CONCLUSION: There are self specific miRNA express spectra and sequence signature of different cells in different tissues, and they can serve as specific molecule marks of some tissues or cells. In the different cell developmental stage, there are different miRNA, and they determine cell differentiation direction and differentiation phase as a switch of timing and direction location differentiation. There are self specific miRNA in various kinds of stem cells, and there are also specific miRNA expressions in different differentiation stage of stem cells. They play a critical role in the maintenance of stem cell self-renewal and multi-directional differentiation characteristics. MiRNA as a new regulator RNA of gene expression provides a new way for the research of stem cells. Related Articles | Metrics

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Directional differentiation of cartilage seed cells in tissue engineereding Du Zhi-po, Bei Kang-sheng 2011, 15 (10):  1879-1883.  doi: 10.3969/j.issn.1673-8225.2011.10.038 Abstract ( 207 )   PDF (642KB) ( 368 )   Save BACKGROUND: Cartilage repair has been an orthopedic therapy problem and a hot research spot. With the development of tissue engineering in recent years, the idea of the application of seed cells induced into cartilage cells and the construction of tissue engineered cartilage repair of cartilage defects has received much attention, which has obtained some achievements. OBJECTIVE: To investigate the selection of seed cells of tissue engineered cartilage, culture condition of chondrogenesis directional induced differentiation, especially for the effects of cell factors, induction methods, culture condition, and other factors. METHODS: The articles in CNKI and PubMed database (2000-01/2010-09) were retrieved by computer. The index words were “chondrification, cartilage defect, stimulating  actor, cartilage repair, tissue engineering” in Chinese and in English, respectively. Literatures were limited to Chinese and English languages. The foundation and clinical research of chondrification, induced differentiation, tissue engineering scaffolds were included, outdated and repetitive researches were excluded. Totally 155 literature were obtained from the computer screen, according to the inclusion and exclusion criteria, 41 documents of them were included finally. RESULTS AND CONCLUSION: Seed cells under the effect of certain cell factors can be directed to differentiate into cartilage cells. It can provide new ideas for successful treatment of articular cartilage defect by selecting appropriate seed cells and cell factors induced into cartilage tissue engineering. Directional cartilage differentiation factor transforming growth factor-β (TGF-β), basic fibroblast growth factor (bFGF), osteogenin, insulin-like growth factor (IGF), growth hormone (GH) were control induced by medium additives, gene transfer, different training methods, and other ways to promote the transformation into cartilage. With the deep investigation addressing induced differentiation of cartilage, this research provides effective basis for the problem of articular cartilage defect repair solved by cartilage tissue engineering in clinical application, and has a wide prospect of clinical application. Related Articles | Metrics

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Treatment of severe aplastic anemia with stem cells from related/unrelated and/or mesenchymal stem cell co-transplantation Yan Hong-min, Wang Zhi-dong, Zhu Ling, Xue Mei, Liu Jing, Pan Shi-ping, Ding Li, Wang Heng-xiang 2011, 15 (10):  1884-1888.  doi: 10.3969/j.issn.1673-8225.2011.10.039 Abstract ( 317 )   PDF (1853KB) ( 411 )   Save BACKGROUND: Hemopoietic stem cell transplantation is the preferred method for the young patients with severe aplastic anemia (SAA), but most patients with SAA without a suitable donor in China. Haploidentical or unrelated hemopoietic stem cell transplantation is still at the exploratory stage at home and abroad, the reports addressing hemopoietic stem cell transplantation combined with mesenchymal stem cell transplantation are rare. OBJECTIVE: To observe the efficacy for SAA with stem cell from related/unrelated and/or hemopoietic stem cell transplantation. METHODS: A total of 10 patients aged 3-52 years with SAA were treated with related HLA-matched (n=2), haploidentical (n=5), unrelated peripheral blood and/or bone marrow hematopoietic stem cell transplantation (n=3), including 5 patients were treated with mesenchymal stem cell co-transplantation. The conditioning regimen mainly included cyclosphosphamide (CY), fludarabine (Flu), and anti-thymocyte globulin (ATG), mycophenolate mofetil, cyclosporin A and short course of methotrexate (MTX) to prevent graft-versus-host disease (GVHD). Patients with haploidentical were treated with myleran and CD25 monoclonal antibody based on the above basis. The conditioning regimen of 5 patients with homogenic is ATG + methylprednisolone. The volume of infusion of mesenchymal stem cells is (0.27-1.85)×106/kg. Patients treated or not treated with mesenchymal stem cell transplantation, transfusion hematopoietic stem cells of them are nucleus cells (7.4-17.38) ×108/kg and (6.09-13.68) ×108/kg, respectively. RESULTS AND CONCLUSION: Except 1 case was a not successful haploidentical transplant patient and died of complications at 36 days, the chromosome and DNA fingerprint detection after transplantation in the remaining patients showed the donors completely received hematopoietic stem cell transplantation. The volume of neutrophil reached 0.5×109 L-1 after transplantation, blood platelets count ≥20×109 L-1, the median time was 12 days and 13 days. The trend of hematopoietic function recovery speed was homogenic transplantation > peripheral blood and/or bone marrow mesenchymal stem cell transplantation > simple peripheral blood and/or bone marrow stem cell transplantation, while the hematopoiesis recovery of the 52-year-old patient with related HLA-matched is the lowest. Case 1 and case 6 with unrelated peripheral blood and/or bone marrow hematopoietic stem cell transplantation suffered gradeⅠacute GVHD, case 2 and case 10 with haploidentical transplantation suffered circumscribed chronic GVHD after gradeⅡ acute GVHD, the quality of life in the remaining patients is good after transplantation, without chronic GVHD. In addition to case 3 suffered severe infection who was not received mesenchymal stem cell transplantation, the remaining patients were not suffered severe infection and hemorrhage after transplantation. The results suggested that hematopoietic stem cells are safe and effective treatment of refractory severe aplastic anemia, which combined with mesenchymal hematopoietic stem cells, rapid hematopoietic recovery and few transplant complications in patients. Related Articles | Metrics

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Autologous hematopoietic stem cell transplantation and allogeneic umbilical cord mesenchymal stem cell transplantation in the treatment of neural degenerative diseases Wang Li-ming, Zhou Jian-jun, Bai Wen, Bai Bing, Li Jian-jun, Wang Li-hua, Liu Yong-jun 2011, 15 (10):  1889-1892.  doi: 10.3969/j.issn.1673-8225.2011.10.040 Abstract ( 327 )   PDF (1643KB) ( 634 )   Save BACKGROUND: Autologous hematopoietic stem cell transplantation (AHSCT) and allogeneic umbilical cord mesenchymal stem cell (UC-MSC) technology are applied in clinic, trying to find a new feasible method of the treatment of neural degenerative disease. OBJECTIVE: To study the feasibility of AHSCT CD34+ and UC-MSC in the treatment of neural degenerative disease. METHODS: A total of 21 patients with neural degenerative disease were selected, including 15 cases with motor neuron disease (MND), 6 cases with spinocerebellar ataxia. A volume of 4 mL UC-MSC or AHSCT solution was injected into subarachnoid space through lumbar puncture; the number of injected cells was 1.0×107 each time. At 3 months following stem cells therapy, all patients were scored. RESULTS AND CONCLUSION: In 15 patients with MND, the total effective rate was 80%. There were significant differences between the acute lateral sclerosis (ALS) functional rating scale and the self-assessment questionnaire scores before and after treatment (P < 0.05). Six cases with spinal cerebellar ataxia underwent international cooperative ataxiarating scale (ICARS). The grade of disease condition was reduced in 4 cases; the total effective rate was 77%. Four of the 21 cases had mild intracranial hypotension headache (after lumbar puncture). Two of the 14 patients with the treatment of UC-MSC had transient fever at 2 hours after treatment. The remaining patients had no significant adverse reactions after treatment. The results primarily demonstrated that the AHSCT and UC-MSC have a positive clinical effect, which are safe and feasible in the treatment of neural degenerative disease. Related Articles | Metrics

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Promoting effect of bone marrow serum on angiogenesis of chick embryo chorioallantoic membrane Tang Yu, Zhong Zhi-ying, Sheng Guo-tai, Ge Yu-zhi 2011, 15 (10):  1893-1896.  doi: 10.3969/j.issn.1673-8225.2011.10.041 Abstract ( 255 )   PDF (313KB) ( 305 )   Save BACKGROUND: Growth factor can promote the development of collateral vessels. The synergistic effects of multiple factors are obvious. Many kinds of growth factors are enriched in bone marrow serum. OBJECTIVE: To determine effects of bone marrow serum on angiogenesis in chick chorioallantoic membranes following vascular intimal injury. METHODS: A total of 70 fertilized chick eggs were incubated at (37.5±0.5) ℃ and made a window in every egg at day 7. The survival chick embryos were randomly divided into six groups: normal saline group, normal blood serum group, normal bone marrow serum group, blood serum of rabbit suffered with vascular intimal injury group, bone marrow serum of rabbit suffered with vascular intimal injury group and vascular endothelial growth factor group (n=10), which were respectively treated with 5 μL rabbit normal blood serum, 5 μL rabbit normal bone marrow serum, 5 μL blood serum of rabbit suffered with vascular intimal injury, 5 μL bone marrow serum of rabbit suffered with vascular intimal injury, 5 μL normal saline and 0.3 μg vascular endothelial growth factor in chick chorioallantoic membranes, for 3 consecutive days. Pictures were taken by a digital camera, and the total number of the vessels around the disc was measured. RESULTS AND CONCLUSION: The total numbers of vessels in normal bone marrow serum group and blood serum of rabbit suffered with vascular intimal injury group were obviously higher than that of normal blood serum group. Large and middle vessels were obviously proliferated. Large and middle vessels were more observed in the blood serum of rabbit suffered with vascular intimal injury group. These indicated that normal bone marrow serum can stimulate angiogenesis in models of chick chorioallantoic membranes. Compared with vascular endothelial growth factor, both the normal bone serum and the bone marrow serum of rabbit suffered from vascular intimal injury were more effective in angiogenesis. The blood serum and bone marrow serum from vascular intimal injured rabbit at day 7 can significantly promote angiogenesis in chick chorioallantoic membranes, which was better than the vascular endothelial growth factor group. Related Articles | Metrics

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Umbilical cord blood mesenchymal stem cells traced and transfected by recombinant lentivirus vector with enhanced green fluorescent protein for treatment of ischemic necrosis of the femoral head in rabbits Li Tao-tao, Wang Xiang-da, Tian Shao-qi, Sun Kang 2011, 15 (10):  1897-1900.  doi: 10.3969/j.issn.1673-8225.2011.10.042 Abstract ( 231 )   PDF (525KB) ( 317 )   Save BACKGROUND: At present, studies concerning human umbilical cord blood mesenchymal stem cells (UCB-MSCs) transplantation for repair of rat spinal cord injury, brain tumor, and myocardial infarction have been reported, and studies that human UCB-MSCs were induced differentiation into osteogenic cells under certain conditions have also been reported at home and abroad. But application of UCB-MSCs transplantation in the treatment of osteonecrosis of animals has not yet been reported. OBJECTIVE: To observe the repair results of recombinant lentivirus vector tracing enhanced green fluorescent protein (EGFP)-transfected UCB-MSCs transplantation in treatment of ischemic necrosis of the femoral head in rabbits. METHODS: Bone morphogenetic protein-2 gene plasmid, recombinant lentivirus vector carrying EGFP and UCB-MSCs were co-cultured. Rabbit models of femoral head defects were made and randomly divided into 3 groups. There was no treatment in the normal group, control group with bone defects and experimental bone defects filled with UCB-MSCs tracing transfected by recombinant lentivirus vector carrying EGFP. At 4 and 8 weeks after treatment, the imaging and histological of the femoral head were observed. RESULTS AND CONCLUSION: Imaging and histology results showed that there were osteogenic response and new bone formation in the experimental group at 4 weeks, and the bone defects were basically repaired at 8 weeks after treatment. In the control group, the bone defects filled with fibrous connective tissue fiber connective tissues at 4 weeks, and the osteosclerosis could be found surrounding femoral head, bone defects filled with fibrous connective tissue fibers and bone trabecula distributed disorderly. The recombinant lentivirus vector tracing EGFP-transfected into UCB-MSCs has strong effects bone conduction and can repair ischemic necrosis of the femoral head. Related Articles | Metrics