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03 September 2014, Volume 18 Issue 37 Previous Issue    Next Issue

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Effect of intermittent tensile stress on cytoskeleton of bone marrow mesenchymal stem cells during osteogenic differentiation in osteoporosis rats Ouyang Ning-juan, Fu Run-qing, Zhang Peng, Wu Yu-qiong, Wang Jie, Jiang Ling-yong, Fang Bing 2014, 18 (37):  5905-5910.  doi: 10.3969/j.issn.2095-4344.2014.37.001 Abstract ( 307 )   PDF (896KB) ( 720 )   Save BACKGROUND: Cytoskeleton plays an important role in the transduction of mechanical signal, and intermittent tensile stress can promote osteogenic differentiation. However, there is no relevant study about the change of cytoskeleton in osteoporosis rat bone marrow mesenchymal stem cells under intermittent tensile stress. OBJECTIVE: To investigate the effects of intermittent tensile stress on the cytoskeleton of osteoporosis rat bone marrow mesenchymal stem cells during osteogenic differentiation.  METHODS: Bone marrow mesenchymal stem cells were obtained from osteoporosis rats and cultured in vitro. The 5%, 10% and 15% tensile stress were strained on the bone marrow mesenchymal stem cells through FX-4000T Flexcell. No stress was in the control group. Osteogenic differentiation of bone marrow mesenchymal stem cells was observed through alkaline phosphatase staining, while the change of cytoskeleton was observed by confocal laser scanning microscopy with figures collected for analysis by Image-ProPlus 6.0 software. The area of cells, ratio of length to width and integrated fluorescence intensity of cytoskeleton protein F-actin were measured. RESULTS AND CONCLUSION: Under tensile stress, bone marrow mesenchymal stem cells from osteoporosis rats arranged in the direction vertical to mechanical stimulation. Cells under different tensile stress differentiated towards osteoblasts. The result of alkaline phosphatase staining showed the most significant difference in 10% group, and quite an amount of cells lining lost succession in the 15% group. Under stress, the F-actin filaments were rearranged in parallel accordingly, which showed a reconstruction of cytoskeleton. Imaging analysis indicated that the area of bone marrow mesenchymal stem cells was decreased in 10% and 15% groups (P < 0.05) with the increased ratio of length to width (P < 0.05), and expression of F-actin increased in5%, 10%, 15% groups (P < 0.05) after tensile stress. Under mechanical stimulation, the cytoskeleton of bone marrow mesenchymal stem cells from osteoporosis rats is shown to have corresponding alterations during osteogenic differentiation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Human nerve growth factor beta-modified bone marrow mesenchymal stem cells from the rabbit mandible by transfection of lentiviral vectors Liu Xiao-chang, Zhao Ying-hua, Yang Zi-gui, Wang Lei 2014, 18 (37):  5911-5915.  doi: 10.3969/j.issn.2095-4344.2014.37.002 Abstract ( 348 )   PDF (697KB) ( 830 )   Save BACKGROUND: Central nerve damage and peripheral nerve injury are common clinical problems that have no ideal treatment. Nerve growth factor has an important role in neuronal repairing and growth. But its local injections may have shorts of inactivation and loss. OBJECTIVE: To construct human nerve growth factor beta recombinant plasmids, which are transfected into bone marrow mesenchymal stem cells from the rabbit mandible by lentiviral vectors, and to investigate the bioactivity of human nerve growth factor beta. METHODS: pDC316-hNGFβ-mCMV-EGFP plasmids were constructed via lentiviral vectors using Hind III+Not I digestion. Bone marrow mesenchymal stem cells from the rabbit mandible were isolated and cultured, and then transfected by recombinant plasmids. The expression of human nerve growth factor beta in transfected cells was detected by ELISA method. RESULTS AND CONCLUSION: pDC316-hNGFβ-mCMV-EGFP plasmids were proved to be constructed successfully by gene sequencing and enzyme identification. The transfected cells under a fluorescence microscope emitted green fluorescence, and the fluorescence intensity had no change with incubation time. The expression of human nerve growth factor beta was maintained at a level of 25 μg/L at 7 days after cell transfection, and the bioacitivty was increased significnalty. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of basic fibroblast growth factor via coronary venous retroperfusion on bone marrow mesenchymal stem cell differentiation in vivo Wang Xiao, Zhen Lei, Miao Huang-tai, Wu Xing-xin, Ren Hong-mei, Shi Shu-tian, Qiao Yan,Liu Xin-min, Que Bin, Nie Shao-ping 2014, 18 (37):  5916-5922.  doi: 10.3969/j.issn.2095-4344.2014.37.003 Abstract ( 318 )   PDF (2530KB) ( 518 )   Save BACKGROUND: In vitro studies have demonstrated that basic fibroblast growth factor (bFGF) promote the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. However, it is unclear whether coronary venous retroperfusion of bFGF stimulates BMSCs differentiation in vivo. OBJECTIVE: To evaluate the effects of coronary venous retroperfusion of bFGF on BMSCs differentiation in vivo. METHODS: BMSCs from 12 dogs were isolated by density gradient centrifugation and expanded in vitro. These cells were transfected by enhanced green fluorescence protein (EGFP) lentiviral vector and the transfection efficiency was analyzed. Acute myocardial infarction was induced by ligation of left anterior descending coronary artery. After 1 week, 10 survival animals were randomized to BMSCs group (n=5) and bFGF+BMSCs group (n=5).bFGF- and EGFP-positive BMSCs were reversely infused via coronary vein using over-the-wire balloon catheter. One week after infusion, the number of EGFP-positive cells co-staining factor VIII and troponin I was compared between the two groups by immunofluorescence method. RESULTS AND CONCLUSION: BMSCs were successfully transfected by EGFP and the transfection efficiency was 85%. Immunofluorescence showed that EGFP-positive BMSCs were observed in 23.5% of slides. There were more EGFP-positive cells co-staining VIII and troponin I in the bFGF+BMSCs group than in the BMSCs group (P < 0.05). Thus, the coronary venous retroperfusion of bFGF enhances the differentiation of BMSCs into vascular endothelial cells and cardiomyocytes. Combined delivery of bFGF and BMSCs can exert a synergistic effect to promote cardiac repair. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Isolation and identification of bone marrow mesenchymal stem cells from transgenic rats with green fluorescent protein gene Wang Yang, Cao Zhi-qiang 2014, 18 (37):  5923-5928.  doi: 10.3969/j.issn.2095-4344.2014.37.004 Abstract ( 333 )   PDF (2763KB) ( 505 )   Save BACKGROUND: Mesenchymal stem cells from transgenic animals carry green fluorescent protein that can be stably expressed in viable cells and can be used to rapidly screen modified cells in comparison with traditional virus and plasmids. OBJECTIVE: To study the biological features of bone marrow derived stromal cells from transgenic rats with green fluorescent protein gene. METHODS: Bone marrows were isolated from the bilateral long bones of 2-week-old transgenic rats with green fluorescent protein gene, and cultivated in conditioned culture medium to obtain bone marrow mesenchymal stem cells with green fluorescent protein gene. The passage 5 bone marrow mesenchymal stem cells with green fluorescent protein gene were analyzed by flow cytometry to detect cell surface molecules, and were induced in calcium or adipogenic induction medium. After calcium induction, the alizarin red staining was performed to test the formation of calcium concentration. After adipogenic induction, bone marrow mesenchymal stem cells were detected with oil red O staining. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells, stably expressing green fluorescent protein gene, were obtained successfully from the transgenic rat with green fluorescent protein gene. The cells expressed CD90, CD105 strongly, but did not express or weakly expressed CD14 andCD45. After 3 weeks of calcium induction, jacinth calcium salts could be detected in the cells by alizarin red staining. At 3 weeks of adipogenic induction, the cells showed positive oil red O staining. The green fluorescent protein gene, as report gene, exerts no effect on the cell surface molecules and multilineage potential of bone marrow mesenchymal stem cells, which can be used as an efficient tracer. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Raman spectroscopy of bone marrow mesenchymal stem cells in medium-frequency pulsed electromagnetic fields Cui Xiang-rong, Su Wei, Wu Zhi-hui, Meng Ling-jing, Huang Zhao, Qin Wan-an 2014, 18 (37):  5929-5934.  doi: 10.3969/j.issn.2095-4344.2014.37.005 Abstract ( 412 )   PDF (1693KB) ( 512 )   Save BACKGROUND: Studies about low-frequency pulsed electromagnetic fields interfering with bone marrow mesenchymal stem cells proliferation and differentiation are many, but the Raman spectra of single stem cells irradiated in electromagnetic fields analyzed by surface Raman spectroscopy analysis are rarely reported. OBJECTIVE: To compare the difference in Raman spectra of bone marrow mesenchymal stem cells with or with no irradiation of 3 000 Hz pulsed electromagnetic fields. METHODS: Bone marrow mesenchymal stem cells isolated from Sprague-Dawley rats were cultured and identified. Passage 3 cells were inoculated into 6-well plates and divided into two groups: pulsed electromagnetic field irradiation group and blank control group. After cultured for 7 days, cells in the two groups were transferred to physiological saline, and 30 cells were randomly collected from each group. Four Raman spectra were harvested from each cell and the average relative intensity of Raman spectra was calculated and compared between two groups. RESULTS AND CONCLUSION: There were the same Raman peaks in the two groups, and the waveforms were basically same in the two group based on the curve mapping by origin 7.0 software. The peak value in the irradiation group was decreased compared with the blank control group. Laser optical tweezers Raman spectroscopy can be applied to study the biochemical changes of a single stem cell at the molecular level. The Raman spectra of bone marrow mesenchymal stem cells irradiated by 3 000 Hz pulsed electromagnetic fields differ from those without irradiation, and the peak also lowered after irradiation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Different pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state Li Xiang-li, Zhang Xiao-ying, Zang Li, Wang Xiao-fang 2014, 18 (37):  5935-5941.  doi: 10.3969/j.issn.2095-4344.2014.37.006 Abstract ( 348 )   PDF (805KB) ( 808 )   Save BACKGROUND: Recent studies have found that bone marrow mesenchymal stem cells play an important role in the development of multiple myeloma, especially for angiogenesis, but whether the pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state is different is yet unclear. OBJECTIVE: To investigate the difference in the pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state. METHODS: Bone marrow samples were extracted from 13 patients with multiple myeloma before treatment and after four therapeutic cycles to isolate bone marrow mesenchymal stem cells. ELISA assay was used to detect levels of vascular endothelial growth factor, hepatocyte growth factor and basic fibroblast growth factor in the supernatant of bone marrow mesenchymal stem cells. The proliferative ability and movement of human umbilical vein endothelial cells cultured in the supernatant of bone marrow mesenchymal stem cells were detected by MTT assay and Transwell assay, respectively. The pro-angiogenesis ability of bone marrow mesenchymal stem cells was measured by Matrigel in vitro. RESULTS AND CONCLUSION: The levels of vascular endothelial growth factor, hepatocyte growth factor and basic fibroblast growth factor in the cell supernatant were significantly higher in active myeloma than those in remissive myeloma (P < 0.05). The supernatant of bone marrow mesenchymal stem cells of active myeloma more significantly promoted the proliferation, migration and blood vessel formation of human umbilical vein endothelial cells than that of remissive myeloma in a dose-dependent manner (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells from active myeloma have stronger pro-angiogenesis ability than those from remissive myeloma. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Meropenem effects on biological characteristics of umbilical cord-derived mesenchymal stem cells during umbilical cord collection Liu Jun-jiang, Zhou Jian-yu, Huang Wen-jing, Hong Jing-xin 2014, 18 (37):  5942-5946.  doi: 10.3969/j.issn.2095-4344.2014.37.007 Abstract ( 429 )   PDF (703KB) ( 501 )   Save BACKGROUND: Due to the difficulty in the control of delivery and collection process, antibiotics are often added into the preservation fluid in order to avoid bacterial contamination but not affect cell growth and proliferation. OBJECTIVE: To observe the effects of meropenem on the proliferation and differentiation potential of umbilical cord-derived mesenchymal stem cells. METHODS: There were two groups in this experiment: control group, preservation fluid with penicillin- streptomycin (final concentration of 100 U/mL); experimental group, preservation fluid with meropenem (final mass concentration of 1.0 mg/L). 100 umbilical cord samples were collected in each group, and the positive rate was calculated. After isolation and culture, the passage 3 cells were used to draw a growth curve, flow cytometry analysis was used for phenotype determination, and osteogenic and adipogenic differentiation of cells were detected. RESULTS AND CONCLUSION: The contamination rates were 3% (3/100) in the experimental group and 20% (20/100) in the control group, indicating that meropenem can obviously reduce the contamination rate. In the experimental group, the morphology, differentiation potential and cell phenotype of the passage 3 cells were all normal. The proliferation ability of cells showed no difference between the two groups. Therefore, meropenem can be added to the preservation fluid. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord blood plasma can replace fetal bovine serum for primary culture, proliferation and cryopreservation of umbilical cord mesenchymal stem cells Wu Jie-ying, Lu Yan, Chen Jin-song, Zhu Lu, Gan Wen-ting 2014, 18 (37):  5947-5954.  doi: 10.3969/j.issn.2095-4344.2014.37.008 Abstract ( 579 )   PDF (3362KB) ( 1212 )   Save BACKGROUND: Fetal bovine serum based media used for expanding and cryopreserving human mesenchymal stem cells raise safety concerns in the clinical setting. OBJECTIVE: To investigate the feasibility of human umbilical cord blood plasma as a replacement for fetal bovine serum in culture and cryopreservation of human mesenchymal stem cells derived from umbilical cord. METHODS: Umbilical cord blood units were suitable for this research if they fulfilled the donor selection criteria of the Guangzhou Cord Blood Bank strictly. Cord blood plasma was ready to use after collected from the plasma reduction during the suitable cord blood units processing and pooling. Umbilical cord mesenchymal stem cells were harvested from the umbilical cord tissue of health full-term newborns after delivery by enzyme digestion and were cultured in the presence of Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing either fetal bovine serum or pooled cord blood plasma. Morphology, proliferation, immunophenotype detected by flow cytometry and differentiation toward adipogenic and osteogenic lineages were utilized for investigating the effect of media on umbilical cord mesenchymal stem cells after 3-5 passages. Then cells were cryopreserved in media containing 10% dimethyl sulfoxide, 20% fetal bovine serum or 20% pooled cord blood plasma for at least 6 months. Viability, adhesion, proliferation, immunophenotype and osteogenic differentiation of the cells were assessed after thawing. RESULTS AND CONCLUSION: The morphology (spindle-shaped and plastic-adherent), phenotype and differentiation potential (osteogenic and adipogenic) were almost indistinguishable between cells cultured in fetal bovine serum or cord blood plasma medium, while cells grown in cord blood plasma medium demonstrated significantly higher proliferation rates than those in medium containing fetal bovine serum. After thawing, the cells maintained their adherence to the culture surface and differentiation potential to osteoblasts, but cells from cord blood plasma cryopreservation medium showed significantly better plastic attachment and produced greater cell numbers than fetal bovine serum for the first three post-thaw passages. The results demonstrate that cord blood plasma can sever as an effective substitute to fetal bovine serum for growth, maintenance and differentiation of umbilical cord mesenchymal stem cells, and thus it will be a safe choice for clinical-scale production of human mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Exosomes derived from human umbilical cord blood mesenchymal stem cells: isolation, identification and biological characteristics Zhang Juan, Liu Feng, Zhang Wei, Cong Xu, Wang Cai-sheng, Wei Lai 2014, 18 (37):  5955-5960.  doi: 10.3969/j.issn.2095-4344.2014.37.009 Abstract ( 893 )   PDF (2698KB) ( 2626 )   Save BACKGROUND: Exosomes are membrane vesicles secreted by mesenchymal stem cells. Increasing studies have shown that mesenchymal stem cells can secrete exosomes via paracrine function to play a role in tissue injury. However, reports on how to isolate and identify exosomes derived from human umbilical cord blood mesenchymal stem cells are few. OBJECTIVE: To extract, purify and identify exosomes derived from human umbilical cord blood mesenchymal stem cells. METHODS: The cell culture supernatant of human umbilical cord blood mesenchymal stem cells was collected. Exosome was extracted and purified with ultrafiltration and gradient centrifugation methods. The morphology of exosome was observed by transmission electronic microscope, and the expressions of CD63, CD81, CD90, CD73, CD105, CD29, and CD166 in exosome of mesenchymal stem cells were analyzed by fluorescent activated cell sorting. RESULTS AND CONCLUSION: Mesenchymal stem cells from human umbilical cord blood secreted exosome  which exhibited elliptic or saucer-like shape and its diameter ranged from 40 to 100 nm with membrane structure. Exosome could express the common surface adhesion molecules CD63, CD81 and the surface adhesion molecules CD90, CD73, CD105, CD29, CD166 of mesenchymal stem cells. These findings indicate that exosome may be secreted by mesenchymal stem cells of human umbilical cord blood, which contains plasma membrane proteins of umbilical cord blood mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Comparison of methods for culturing the mesenchymal stem cells from human umbilical cord blood Huang Hong-yu, Liu Guo-ping, Duan Li, Chen Yun-fang, Xiong Jian-yi, Wang Da-ping 2014, 18 (37):  5961-5966.  doi: 10.3969/j.issn.2095-4344.2014.37.010 Abstract ( 461 )   PDF (1732KB) ( 712 )   Save BACKGROUND: The umbilical cord blood is rich of mesenchymal stem cells, which can be used as a new source of seed cells in tissue engineering. OBJECTIVE: To compare two methods for culturing, expanding and purifying the mesenchymal stem cells in vitro isolated from the human umbilical cord blood. METHODS: The full-term birth cord blood of 40 cases was collected under sterile conditions with heparin anticoagulation. Ficoll density gradient centrifugation was used to isolate the mononuclear cells from the umbilical cord blood. The cases were randomly divided into two groups according to the different culture media. Twenty cases of umbilical cord blood were cultured in MesenGro human mesenchymal stem cells culture medium (group A), and the remaining 20 cases of umbilical cord blood were cultured in Dulbecco’s modified Eagle’s medium(group B). The occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and the number of primary cells in the two groups were compared. Cells which grew well were selected to detect the surface markers by flow cytometry. RESULTS AND CONCLUSION: The mean occurrence time of fusiform mesenchymal stem cells and cell colony, culture time and number of primary cells in group A were better than those in group B (P < 0.01). The strong expression of the surface markers of mesenchymal stem cells (CD73 and CD105) was found by flow cytometry, of which the positive rate was 99.1%. No expression of the surface markers of hematopoietic stem cells (CD45 and CD34) was seen, of which the negative rate was 99.3%. The number, morphology, growth rate and culture time of umbilical cord blood mesenchymal stem cells cultured in MesenGro human mesenchymal stem cells culture medium were better than those cultured in Dulbecco’s modified Eagle’s medium. Cells cultured in MesenGro human mesenchymal stem cell culture medium can better express surface markers of mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of SIRT6/NF-kappa B signal axis during hematopoietic stem and progenitor cell senescence induced by tert-butylhydroperoxide in vitro Zhou Yue, Wang Ya-ping, Wang Jian-wei, Ding Ji-chao, Yang Yong-qin 2014, 18 (37):  5967-5971.  doi: 10.3969/j.issn.2095-4344.2014.37.011 Abstract ( 397 )   PDF (658KB) ( 993 )   Save BACKGROUND: SIRT6/NF-κB is the important signal axis to cell senescence, but the effect of SIRT6/NF-κB signal axis to hematopoietic stem and progenitor cell (HSC/HPC) senescence is unclear. OBJECTIVE: To induce (t-BHP) in vitro and to investigate the role of SIRT6/NF-κB signal axis in Sca-1+ HSC/HPC senescence induced by tert-butylhydroperoxide in vitro. METHODS: Sca-1+HSC/HPC was isolated and purified by magnetic activated cell sorting. Sca-1+ HSC/HPC senescence was induced by 100 μmol/L tert-butylhydroperoxide in vitro. The senescence-associated β-Galactosidase staining, cell cycle analysis and culture of mixed hematopoietic progenitor cell were used to investigate the biological effects of tert-butylhydroperoxide on Sca-1+ HSC/HPC senescence. The expression of senescence associated SIRT6, NF-κB mRNA and protein was examined by real-time fluorescence quantitative PCR and western blot assay. RESULTS AND CONCLUSION: Compared with control group, the percentage of positive cells expressing SA-β-Gal and cells in G0/G1 phase increased and the number of forming colony of mixed hematopoietic progenitor decreased in the aging group. It showed lower expression of SIRT6 and higher expression of NF-κB in the aging group. The SIRT6/NF-κB signal axis may play a key role in the Sca-1+ HSC /HPC senescence inducted by tert-butylhydroperoxide. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of induction therapy with bone mesenchymal stem cells on adenosine triphosphate levels in CD4+ T cells determined by ImmuKnow assay in patients receiving renal transplantation Chen Shu-shang, Cai Jin-quan, Wu Cheng-yao, Deng Zhen, Zhu Ling-feng, Zhou Hao, Wang Qing-hua, Tan Jian-ming 2014, 18 (37):  5972-5976.  doi: 10.3969/j.issn.2095-4344.2014.37.012 Abstract ( 326 )   PDF (673KB) ( 540 )   Save BACKGROUND: Bone mesenchymal stem cells have immunological regulation function both in vitro and in vivo, while the effect of bone marrow mesenchymal stem cells on CD4+ T cell immune function in patients receiving kidney transplantation remains unclear. OBJECTIVE: To explore the monitoring significance of CD4+ T-cell immune function by ImmuKnow assay and to determine the effect of induction therapy with bone marrow mesenchymal stem cells on cell immune function in patients receiving kidney transplantation. METHODS: From January 2011 to June 2013, 24 patients receiving allograft renal transplantation with autologous bone marrow mesenchymal stem cells were included and another 48 patients receiving allograft renal transplantation and Simulect induction therapy with various matched preoperative characters served as controls. In both groups, adenosine triphosphate levels in CD4+ T cells in the peripheral blood were determined by the ImmuKnow assay preoperatively and at 14, 30, 60, 90, 180 days postoperatively, as well as during acute rejection and infection episodes. RESULTS AND CONCLUSION: During the 180 days postoperatively, fewer patients in the bone marrow mesenchymal stem cell group had acute rejection and injection than the Simulect group, but no significant differences were observed. Postoperative adenosine triphosphate levels in CD4+ T cells were significantly lower than those determined preoperatively in both groups (P < 0.05), while no significant differences were observed between the two groups. A total of 12 patients in the bone marrow mesenchymal stem cell group and 26 patients in the Simulect group had infection episodes, and the adenosine triphosphate levels in CD4+ T cells during the infection episodes were lower than clinical stable patients in both groups (P < 0.01). For patients receiving renal transplantation, induction therapy with bone marrow mesenchymal stem cells can effectively decrease the cell immune function, which can be reflected by the adenosine triphosphate levels in CD4+ T cells in the peripheral blood determined by the ImmuKnow assay. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells protect against renal ischemia-reperfusion injury through immune regulatory mechanism Hu Hong-lin, Zou Cong, Xi Xiao-qing, Ye Zhen-feng, Jiang Wei 2014, 18 (37):  5977-5982.  doi: 10.3969/j.issn.2095-4344.2014.37.013 Abstract ( 321 )   PDF (750KB) ( 769 )   Save BACKGROUND: Stem cell therapy for renal ischemia-reperfusion injury has been the hot topics for many scholars. Its mechanism is very complex, which could not be explained by simple mechanism of stem cells differentiation. It is the result involving a variety of mechanisms. OBJECTIVE: To investigate the influence on immune cells during the bone marrow mesenchymal stem cell therapy for renal ischemia-reperfusion injury, then to preliminary summarize the immune regulation mechanism of bone marrow mesenchymal stem cell therapy for renal ischemia-reperfusion injury. METHODS: First, we established a model of renal ischemia-reperfusion injury in rats and, cultured and purified rat bone marrow mesenchymal stem cells in vitro. Then, the bone marrow mesenchymal stem cells were transplanted into the rat models. Using flow cytometry detection technology, we analyzed the proportion of CD4+CD25+regulatory T cells of rat spleen cells, discussed the effects on immune cells during the bone marrow mesenchymal stem cell therapy for renal ischemia-reperfusion injury, and then transferred the rat’s spleen cells to the nude mice which were subjected to renal renal ischemia-reperfusion injury. Renal function and renal histological changes of nude mice were assessed. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cell transplantation could significantly inhibit the decrease of CD4+CD25+ regulatory T cell of spleen cells in rats with renal ischemia-reperfusion injury. The transplantation of spleen cells from the above-mentioned rats to nude mice could obviously protect nude mice from renal ischemia-reperfusion injury, characterized by lower serum creatinine, blood urea nitrogen and renal tubule pathologic damage score. Therefore, bone marrow mesenchymal stem cells have protective effects on renal ischemia-reperfusion injury by regulating the immune system. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Allogeneic bone marrow mesenchymal stem cell transplantation for acute myocardial infarction in rats Liang Li-ling, Yang Ting-shu, Li Ping, Feng Bin, Han Bao-shi 2014, 18 (37):  5983-5987.  doi: 10.3969/j.issn.2095-4344.2014.37.014 Abstract ( 341 )   PDF (764KB) ( 663 )   Save BACKGROUND: A number of studies have shown that bone marrow mesenchymal stem cells can survive in the infarcted myocardium and improve cardiac function. OBJECTIVE: To investigate the effects of allogeneic rat bone marrow mesenchymal stem cells on heart failure in acute myocardial infarction models of rats and possible mechanisms. METHODS: Rat bone marrow mesenchymal stem cells were isolated from the bone marrow of 39 male Wistar rats by density gradient centrifugation with Percoll. After ligating anterior descending coronary artery, 39 female Wistar rats were randomly divided into three groups: control group (Dulbecco’s modified Eagle’s medium, n=12), mesenchyma stem cells group (n=15) and mononuclear cells group (n=12). Eight weeks later, hemodynamics and left ventricular function were measured. Histopathological and immunohistochemical examinations were performed. RESULTS AND CONCLUSION: Compared with the control group, left ventricular end diastolic pressure, left ventricular relative weight, the collagen volume fraction of type I and type III in the infarction zone of the left ventricle were all significantly decreased, in contrast to ±dp/dtmax, -dp/dtmax/left ventricular systolic pressure, body weight and vascular density in infarction zone were all significantly increased both in mesenchymal stem cells group and mononuclear cells group. There were no significant differences between two treatment groups except for interventricular septal thickness and vascular density in non-infarction zone. 5-Bromo-2'-deoxyuridine positive cells were observed in the infarction area of mesenchyma stem cells group but no positive cells in mononuclear cells group. Some ball-like cell masses were found positively stained with desmin and cardiac troponin T. Results have suggested that embedded bone marrow mesenchymal stem cells survived in exogenous host hearts. The therapy of mononuclear cells and mesenchymal stem cells could limit the left ventricular remodeling after acute myocardial infarction and improve left ventricular function through angiogenesis inducing and collagen deposition decreasing. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Transplantation of bone marrow mesenchymal stem cells transfected by vascular endothelial growth factor gene promotes foot wound healing in diabetic rats Cai Qian, Wan Jiang-bo, Liang Wen-jia, Liu Yi 2014, 18 (37):  5988-5992.  doi: 10.3969/j.issn.2095-4344.2014.37.015 Abstract ( 341 )   PDF (704KB) ( 772 )   Save BACKGROUND: Diabetic foot ulcers threaten the patients’ health and even survival seriously. It is an international difficult problem and lacks an effective treatment. But gene therapy and stem cell therapy possess special advantages and potential in wound healing. OBJECTIVE: To assess the therapeutic effect of transplantation of bone marrow mesenchymal stem cells transfected by human vascular endothelial growth factor 165 (hVEGF165) gene on foot wound healing in diabetic rats. METHODS: Recombinant adenovirus was established in vitro which expressed hVEGF165 gene and transfected into the third generation of bone marrow mesenchymal stem cells. Totally 120 male Wistar rats were divided into five groups: group A (non-diabetic controls), group B (diabetic controls), group C (Ad-hVEGF165 therapy), group D (stem cell therapy) and group E (transplantation of bone marrow mesenchymal stem cells transfected by Ad-hVEGF165 gene). Rats in the latter four groups were intraperitoneally injected with streptozotocin to induce diabetic models. In all rats, a 3 mm×7 mm rectangular full-thickness skin sample was cut from the instep of the hind foot to make a model of foot wound. The rats were subcutaneously injected at equidistant six points 5 mm distal to the wound edge on the dorsum of the foot: 50 μL PBS per point for group A, 50 μL adenovirus suspension (1×1013 pfu/L) per point for group C, 50 μL stem cell suspension (1×1010/L) per point for group D, and 50 μL adenovirus suspension+50 μL stem cell suspension per point for group E. RESULTS AND CONCLUSION: After injection, the rate of wound healing, the expression of VEGF and the qualities of capillaries in group E were higher when compared with groups B, C, D (P < 0.05), but were lower than those in group A (P < 0.05). Transplantation of bone marrow mesenchymal stem cells transfected by hVEGF165 gene can promote foot wound healing, angiogenesis and expression of VEGF in diabetic rats. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord mesenchymal stem cells for repair of combined radiation-wound skin injury and tumorigenicity in vitro Su Zhong-yi, Yang Zai-liang, Tang Yong-yong, Hu Jiang-wei, Sheng Hong-xia, Xu Man, Zhang Bin, Chen Hu 2014, 18 (37):  5993-5997.  doi: 10.3969/j.issn.2095-4344.2014.37.016 Abstract ( 313 )   PDF (656KB) ( 726 )   Save BACKGROUND: Many scholars have experimentally confirmed the obvious effect of mesenchymal stem cells to repair radiation injury. OBJECTIVE: To preliminarily investigate the mechanism of human umbilical cord mesenchymal stem cells promoting the healing of combined radiation-wound skin injury and whether they possess tumorigenicity in vitro. METHODS: Fifteen Sprague-Dawley rats were randomly divided into three groups, five rats in each group. The right buttock of rats (2.5 cm×2.0 cm) was irradiated with 40 Gy β-rays produced by a linear accelerator, in which a round wound with a diameter of 1.5 cm was made. After 12 hours of modeling, human umbilical cord mesenchymal stem cells at three concentrations (5.0×106, 1.0×107 and 2.0×107) were injected through tail vein of rats, and luciferin (20 mg/kg) was injected intraperitoneally. Cell distribution in vivo was traced using IVIS in vivo imaging system. The ability of human umbilical cord mesenchymal stem cells to form colonies was observed using the colony formation assay with soft agar. RESULTS AND CONCLUSION: Human umbilical cord mesenchymal stem cells injected through tail vein of rats were mostly gathered in the lungs. Cells were accumulated in the injured site of rats injected with 2.0×107 human umbilical cord mesenchymal stem cells; however, the fluorescence signal was not observed in the injured site of rats injected with 5.0×106 and 1.0×107 human umbilical cord mesenchymal stem cells. The other results indicated human umbilical cord mesenchymal stem cells of low dose, medium dose and high dose had no colony formation on soft agar, but the tumor cells had a great ability to form colony. These findings indicate that human umbilical cord mesenchymal stem cells promote healing combined radiation-wound skin injury by local migration and exhibit no tumorigenicity in vitro in a short period. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Adipose-derived mesenchymal stem cells differentiate into inner ear hair cells in guinea pigs induced by progressive addition of cytokines Wang Xiao-yan, Li Bing-bing, Zhang En-feng, Bi Xiao-juan, Liu Li-zhong 2014, 18 (37):  5998-6002.  doi: 10.3969/j.issn.2095-4344.2014.37.017 Abstract ( 254 )   PDF (1606KB) ( 506 )   Save BACKGROUND: Sensorineural hearing loss is mainly caused by missing or damaged hair cells in the inner ear. Application of adipose-derived mesenchymal stem cells to regenerate inner ear hair cells is an effective treatment for hearing loss. OBJECTIVE: To explore the feasibility of in vitro inducing adipose-derived mesenchymal stem cells to differentiate into inner ear hair cell-like cells in guinea pigs. METHODS: Adipose-derived mesenchymal stem cells from guinea pigs were isolated and cultured to the 3rd generation. Cell phenotype was detected using flow cytometry. Cytokines were added for induction and differentiation by stages, including epidermal growth factor, basic fibroblast growth factor, insulin-like growth factor-1, all-trans retinoic acid, brain-derived neurotrophic factor, neurotrophin 3. RESULTS AND CONCLUSION: Adipose-derived mesenchymal stem cells of guinea pigs cultured in vitro were fusiform and showed a swirled adherent growth. Passage 3 cells were positive for CDCD29 and CD44, but negative for CD34 and CD45. After induction, the cells were positive for nestin and GFAP positive at early stage;after 10-day continuous induction, the cells expressed Myosin VIIa and Math1, specific markers of hair cells, indicating that cytokines can directly induce adipose-derived mesenchymal stem cells to differentiating into inner ear hair cells in guinea pigs. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Co-culture of adipose-derived stem cells and osteoblasts under different conditions Zhang Yang, Liu Da-cheng, Yang Xiao-ning 2014, 18 (37):  6003-6007.  doi: 10.3969/j.issn.2095-4344.2014.37.018 Abstract ( 330 )   PDF (2408KB) ( 525 )   Save BACKGROUND: After co-culture with osteoblasts, bone marrow stem cells can be induced to differentiate into osteoblasts. Whether adipose-derived stem cells co-cultured with osteoblasts can differentiate into osteoblasts or not? OBJECTIVE: To observe whether adipose-derived stem cells co-cultured with osteoblasts can differentiate into osteoblasts. METHODS: Adipose-derived stem cells and osteoblasts were isolated from New Zealand white rabbits. Then, passage 3 adipose-derived stem cells were co-cultured with passage 2 osteoblasts in 10% or 5% fetal bovine serum for 14 days. RESULTS AND CONCLUSION: After 7 days of co-culture, some adipose-derived stem cells became round in the two groups. After 14 days of co-culture, adipose-derived stem cells highly differentiated and differentiated cells were similar to mature osteoblasts that were positive for alkaline phosphatase staining and alizarin red staining. The mRNA expression of type I collagen and osteocalcin increased in both two group, especially in the 10% fetal bovine serum group. These findings indicate that adipose-derived stem cells co-cultured with osteoblasts can differentiate into osteoblasts induced by high-concentration serum culture. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Osteogenic and adipogenic differentiation of rabbit adipose-derived mesenchymal stem cells in vitro Lee Soo-min, Wu Zi-zheng, Wang Zhe, Li Zhi, Zhang Jian 2014, 18 (37):  6008-6013.  doi: 10.3969/j.issn.2095-4344.2014.37.019 Abstract ( 366 )   PDF (2325KB) ( 752 )   Save BACKGROUND: Adipose-derived mesenchymal stem cells have the ability to self-renew and have pluripotent potential under specific conditions in vitro, which have broad application prospects in clinical practice. However, isolation and culture of adipose-derived mesenchymal stem cells still appear to have many difficulties and shortcomings. OBJECTIVE: To isolate and culture rabbit adipose-derived mesenchymal stem cells in vitro in order to study their morphology, cell surface markers and biological properties as well as to investigate the osteogenic and adipogenic potentials of adipose-derived mesenchymal stem cells in vitro. METHODS: Primary adipose-derived mesenchymal stem cells were isolated from the subcutaneous adipose tissue of posterior cervical region from New Zealand white rabbits and digested by 0.1% collagenase I. The cells were passaged and amplified by the trypsin digestion. The passage 4 adipose-derived mesenchymal stem cells were induced to differentiate after exposure to adipogenic or osteogenic medium. The oil red O staining, alkaline phosphatase and alizarin red staining were used to detect the results. The cell viability was detected by the cell counting kit 8 method to drawn the growth curve. Cell surface markers were examined using flow cytometry. RESULTS AND CONCLUSION: The adipose-derived mesenchymal stem cells isolated from the subcutaneous adipose tissue of rabbits exhibited a fusiform adherent growth in a vortex pattern, and had a strong capability of proliferation that could be passaged stably to the 10th generation. Flow cytometry results showed that the cells highly expressed CD29, CD90, CD44, but lowly expressed CD45 and CD34. After adipogenic induction, the adipose-derived mesenchymal stem cells were positive for oil red O staining; after osteogenic induction, the cells were both positive for alkaline phosphatase and alizarin red staining. These findings suggest that the adipose-derived mesenchymal stem cells were successfully isolated and cultured from the subcutaneous adipose tissue of rabbits, and these cells are pluripotent with the potential to differentiate into adipocytes and osteoblasts, which are expected to be ideal seed cells for bone tissue engineering. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Research about panaxtrial saponins on the relationship between cerebral ischemic tolerance and proliferation of endogenous neural stem cells Jiang Xiao-feng, Zhang Jie-wen, Luo Zu-ming 2014, 18 (37):  6014-6018.  doi: 10.3969/j.issn.2095-4344.2014.37.020 Abstract ( 396 )   PDF (604KB) ( 725 )   Save BACKGROUND: Cerebral ischemia tolerance can promote proliferation of autologous neural stem cells in the hippocampus of cerebral infarction rats, but panaxtrial saponins effects on the proliferation of autologous neural stem cells in the brain have not been reported.  OBJECTIVE: To explore the relationship of panaxtrial saponins, ischemic preconditioning and proliferation of endogenous neural stem cells in the hippocampus of rats at 7 days after cerebral infarction, and to observe the effect on neurobehavioral scores of rats after cerebral infarction. METHODS: Fifty Sprague-Dawley rats were included and randomly divided into five groups: sham group, ischemia group, ischemic control group, ischemic preconditioning group, and panaxtrial saponins group. In the latter four groups, acute models of cerebral infarction were established using Zea-Longa method. In the sham group, only an incision was made on the neck. The focal-focal ischemic tolerance models were established with twice suture method in the ischemic preconditioning and panaxtrial saponins groups. Sham operation was instead of ischemic preconditioning in the ischemic control group. In the panaxtrial saponins group, rats were given intraperitoneal injection of 100 mg/kg panaxtrial saponins at 7 days before modeling. RESULTS AND CONCLUSION: After 7 days of cerebral infarction, the neurobehavioral score and the number of neural stem cells in the hippocampus were significantly increased in the ischemia group (P < 0.01); compared with the ischemia group, the neurobehavioral scores were lowered in the ischemic preconditioning and panaxtrial saponins groups (P < 0.01), while the number of neural stem cells in the hippocampus was increased (P < 0.01). However, there was no difference between the ischemic preconditioning and panaxtrial saponins groups (P > 0.05). In addition, differences in the neurobehavioral scores and the number of neural stem cells in the hippocampus were insignificant between the ischemic control group and ischemia group (P > 0.05). These findings indicate that panaxtrial saponins can play a role similar to ischemic tolerance, and thus improve neurologic impairment in rats with cerebral infarction. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Salidroside induces the differentiation of mouse bone marrow mesenchymal stem cells into neuron-like cells mediated by calcium/calmodulin signaling pathway Zhao Ling, Zhao Hong-bin, Pan Qian, Li Gen, Wang Jiu-na, Tang Jun-jie 2014, 18 (37):  6019-6023.  doi: 10.3969/j.issn.2095-4344.2014.37.021 Abstract ( 320 )   PDF (753KB) ( 526 )   Save BACKGROUND: Our previous studies have shown that salidroside can induce bone marrow mesenchymal stem cells directly into neuron-like cells, and Ca2+ signal is one important way to achieve its biological signal transduction. OBJECTIVE: To investigate the role and mechanism of the calcium/calmodulin (Ca2+/CaM) signaling pathway inducing bone marrow mesenchymal stem cells to differentiate into nerve cells. METHODS: Bone marrow mesenchymal stem cells were divided into two groups: control groups and salidroside groups. Salidroside groups were induced with different concentrations of salidroside (5, 10, 20, 50 and 100 mg/L) for 24 hours and 100 mg/L salidroside was added to culture cells for different time (12, 24, 48 and 72 hours). Western blot assay was used to detect the expression levels of neural cell marker, microtubule-associated protein 2, and the important protein of Ca2+/CaM signaling pathway: CaM and calmodulin dependent kinase II (CaMK II). Then Ca2+/CaM signaling pathway specific blockers were applied to cells respectively for 30 minutes, including 500 µmol/L EGTA (Ca2+ chelator), 1 mmol/L Nifedipine(L-type Ca2+ channel blocker) and 10 mmol/L LY294002 (PI3K inhibitor). Then, 100 mg/L salidroside was added and cultured for 24 hours. Western blot assay was used to detect the expression of neuron-specific enolase and CaM in the Ca2+/CaM signaling pathway. RESULTS AND CONCLUSION: (1) After inducing with salidroside, the expression of microtubule-associated protein 2 were upregulated (P < 0.01), indicating that salidrosid can induce the neuronal differentiation of bone marrow mesenchymal stem cells. (2) After different concentrations of salidrosid induced bone marrow mesenchymal stem cells for 24 hours, the expressions of CaM and CaMK II were significantly upregulated in the 10 mg/L group ( P < 0.01); For the 100 mg/L salidrosid that was added for cell induction for different time, the expressions of CaM and CaMK II were significantly downregulated in 72-hour group (P < 0.01). (3) After blocking extracellular Ca2+ and PI3K signaling pathway, the expressions of neuron-specific enolase and CaM were higher than those in salidrosid groups (P < 0.05). These results suggest that salidrosid can induce bone marrow mesenchymal stem cell to directly differentiate into nerve cells by inhibiting the Ca2+/CaM signaling pathway. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Amniotic cells protect and repair mouse brain cells following ischemia-reperfusion injury Zheng Yan-tao, Liu Bin, Robert Lodato, Li Qi-lin, Lan Di-hui, Hong Xiao-ying, Xian Hua 2014, 18 (37):  6024-6028.  doi: 10.3969/j.issn.2095-4344.2014.37.022 Abstract ( 315 )   PDF (969KB) ( 931 )   Save BACKGROUND: Amniotic cells are mainly composed of amniotic epithelial cells and amniotic mesenchymal cells, which have multi-differentiation potential and can be transformed into neurons as well as synthesize and release biologically active substances and neurotrophic factors. In preliminary studies, amniotic cells that are transplanted into the brain can significantly promote the regeneration of brain neurons. OBJECTIVE: To explore the role of amniotic cells in mouse brain cells after ischemia-reperfusion injury. METHODS: The model of cerebral ischemia-reperfusion injury was established in Babl/c mice using occlusion of bilateral common carotid arteries, and then brain cells were separated from mice. Amniotic cells were isolated from mouse placenta. Brain cells from Balb/C mice co-cultured with amniotic cells served as experimental group, and brain cells cultured with PBS as control group. RESULTS AND CONCLUSION: The viability of brain cells in the experimental group was significantly higher than that in the control group (P < 0.05). There was no difference in necrotic rate of brain cells between the experimental and control groups after 24 and 72 hours co-culture (P > 0.05); after 48 hours co-culture, however, the necrotic rate of brain cells was significantly lower in the experimental group than the control group (P < 0.05). In cell cycle, the experiment group showed increased S phase cells; while, the control group exhibited increased G1 phase cells and decreased S phase cells. G2 phase cells had no changes in number in both two groups. Through the above results, amnion cells can be proved to protect and promote the regeneration of brain cells of Balb/C mice with ischemia-reperfusion injury, and inhibit cell necrosis and apoptosis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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More than 3 ku proteins in chicken egg extract up-regulate expression of pluripotent genes Oct-3/4 and Nanog Ruan Guang-ping, Yao Xiang, Liu Ju-fen, Shu Fan, Wang Jin-xiang, He Jie, Yang Jian-yong, Zhao Jing, Pang Rong-qing, Pan Xing-hua 2014, 18 (37):  6029-6033.  doi: 10.3969/j.issn.2095-4344.2014.37.023 Abstract ( 315 )   PDF (663KB) ( 745 )   Save BACKGROUND: Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE: To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS: The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per well, total 500 μL. In the control group, 500 μL culture medium was added; in the other three groups, 500 μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION: By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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The relationship of Slit2 and bone marrow mesenchymal stem cells with the angiogenesis Jiang Lai, Zhang Jin-ning, Chai Yuan, Li Fu-chun, Qu Yan-ping, Ma Xue-ling 2014, 18 (37):  6034-6039.  doi: 10.3969/j.issn.2095-4344.2014.37.024 Abstract ( 310 )   PDF (606KB) ( 467 )   Save BACKGROUND: Bone marrow mesenchyme stem cells are important non-hematopoietic stem cells in the bone marrow, which can stimulate angiogenesis. While, Slit can also stimulate angiogenesis, as many studies have proved. OBJECTIVE: To review the biological functions, clinical application and effects of bone marrow mesenchymal stem cells and Slit2 on promoting angiogenesis. METHODS: A computer-based online research of CNKI and PubMed databases was performed to collect articles published between 1980 and 2014 with the keywords “MSCs” and “Slit2” in Chinese and English. There were 436 articles after the initial survey. Finally, 65 articles were included according inclusion and exclusion criteria. RESULTS AND CONCLUSION: Both bone marrow mesenchymal stem cells and Slit2 play an important role in promoting angiogenesis, but the relevance of bone marrow mesenchymal stem cells and Slit2 is still controversial. If assuming that bone marrow mesenchymal stem cells secrete Slit2, more researches should be done to reveal whether bone marrow mesenchymal stem cells promoting angiogenesis is relevant to Slit2 and through which signaling pathway Slit2/Robo functions to adjust bone marrow mesenchymal stem cells thus to promote angiogenesis. If relevant, the transplantation of the Slit2 and bone marrow mesenchymal stem cells will be a promising treatment of cerebral infarction and other central nervous injuries. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Hedgehog signal regulates the chondrogenesis from bone marrow mesenchymal stem cells: controlling methods and cross-talking relationship with other signals need further studies Liu Kuan, Wu Xing 2014, 18 (37):  6040-6045.  doi: 10.3969/j.issn.2095-4344.2014.37.025 Abstract ( 403 )   PDF (876KB) ( 510 )   Save BACKGROUND: The hedgehog pathway has paid an important role in the progress of chondrogenesis from bone marrow mesenchymal stem cells. However, the definite signal transduction pathway and cross-talking relationship with other common signal pathways are still poorly understood and the researches related to this field is to continue as a hotspot in the future study. OBJECTIVE: To investigate the research progress of hedgehog signal pathway on the regulation of the chondrogenesis from bone marrow mesenchymal stem cells and the relationship between hedgehog and other signal pathways in the process. METHODS: A computer-based online search in CNKI, PubMed and Google Scholar databases was performed using key words of “Hedgehog, IHH, SHH, bone marrow mesenchymal stem cells, cartilage, chondrogenesis” in English and Chinese, respectively. Literatures related to the process of chondrogenesis from bone marrow mesenchymal stem cells were included and 36 articles were extensively summarized for review. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells are currently accepted optimal cell seeds for the cartilage tissue engineering, and hedgehog is a critical signal molecule in the development of skeletal system. The IHH and SHH in hedgehog signal closely participate in controlling the processes of bone marrow mesenchymal stem cell proliferation and chondrogenesis, chondrocyte phenotype maintenance and cooperation with other common single pathways. However, the specific signal transduction mechanism and cross-talking contact with other signal pathways still need to be further studied, and it stands for the future research directions. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Intranasal delivery of bone marrow mesenchymal stem cells for brain injuries: how many questions to be verified? Yan Xue-jing, Wang Xin-ling, Yang Mi-mi, Hou Wei-jian 2014, 18 (37):  6046-6050.  doi: 10.3969/j.issn.2095-4344.2014.37.026 Abstract ( 323 )   PDF (577KB) ( 460 )   Save BACKGROUND: Transplantation of bone marrow mesenchymal stem cells can promote repair of brain injuries in animals. OBJECTIVE: To summarize the research progress in intranasal delivery of bone marrow mesenchymal stem cells to the brain. METHODS: A computer-based online retrieval of PubMed and Wanfang databases was performed to search papers published during January 1999 to January 2014 with the key words of “bone marrow mesenchymal stem cells, brain injury, transplantation” in English and Chinese. Thirty-eight papers were included in the final analysis. RESULTS AND CONCLUSION: Nowadays, many studies have been certified that the transplantation of bone marrow mesenchymal stem cells can significantly ameliorate the function of cranial nerve in animal models of brain injury. Many researchers have searched for the transplantation methods and approaches and have made progresses in many aspects. In this article, we compare the different transplantation ways of bone marrow mesenchymal stem cells to the brain. We focus on the intranasal transplantation route in the following aspects: processing of the nasal mucosa; delivery route to the brain; labeling and intracranial observation of stem cells; animal experiments. We conclude that the intranasal delivery of bone marrow mesenchymal stem cells to the brain has a wide clinical application as a noninvasive transplantation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Odontoblastic differentiation of dental pulp cells in vitro: odontogenic stimuli and the underlying mechanisms Zou Hui-ru, Steven Brookes, Yang Xue-bin 2014, 18 (37):  6051-608.  doi: 10.3969/j.issn.2095-4344.2014.37.027 Abstract ( 438 )   PDF (628KB) ( 765 )   Save BACKGROUND: Studies have shown that dental pulp cells can differentiate into odontoblastic like cells under certain stimulations, which is regulated by several signal pathways network. OBJECTIVE: To summarize the research progress of stimulating molecules and underlying molecular mechanisms of dental pulp cells’ odontoblastic differentiation.  METHODS: A computer-based online search of CNKI and Wanfang databases was undertaken for the related Chinese articles dated from January 1998 to July 2014 with the keywords of “dental pulp cells” and “odontoblastic differentiation” in Chinese. Meanwhile, PubMed database was searched for the related English articles dated from January 1998 to July 2014 with the same keywords in English. Those with unrelated research subjects, or repetitive studies were excluded. Finally, 63 articles were reviewed. RESULTS AND CONCLUSION: Various stimuli, including bone morphogenetic protein, β-glycerophosphate and ascorbic acid, can stimulate dental pulp cells differentiation into odontoblastic like cells under certain conditions regulated by several signal pathways network. But odontoblastic differentiation is a complex process, and further research into stimulating molecules and underlying molecular mechanisms of dental pulp cells’ odontoblastic differentiation is significantly important for clinical applications of dental regenerative therapy. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell transplantation for spinal cord injury: prospects and issues Li Mai, Ao Li-juan 2014, 18 (37):  6059-6063.  doi: 10.3969/j.issn.2095-4344.2014.37.028 Abstract ( 799 )   PDF (568KB) ( 469 )   Save BACKGROUND: At present, spinal cord injury treatment is still the worldwide difficult problem. Using the method of stem cells transplantation to treat the spinal cord injury is one of the hotspots of spinal cord injury repair research in recent years. OBJECTIVE: To summarize the progress and application prospects of stem cell transplantation in the treatment of spinal cord injury. METHODS: A computer-based search of PubMed and CNKI was retrieved for relevant articles concerning stem cell transplantation for treatment of spinal cord injury published after 2000. The keywords were “spinal cord injury, stem cell, cell therapy” in English and Chinese, respectively. Meta-analysis and secondary literature were excluded as well as repetitive or old literature. Finally, 52 articles were included in result analysis. RESULTS AND CONCLUSION: This article summarizes types and biological characteristics of stem cells, basic mechanism, techniques and therapeutic efficacy of stem cell transplantation in the treatment of spinal cord injury, and proposes the issues and prospects concerning the stem cells transplantation for treatment of spinal cord injury. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Cardiac stem cells in cardiac tissue engineering: present and future Li Run-qin, Huang Chun 2014, 18 (37):  6064-6068.  doi: 10.3969/j.issn.2095-4344.2014.37.029 Abstract ( 272 )   PDF (587KB) ( 458 )   Save BACKGROUND: For a long time, the myocardium of adult mammalians is the terminally differentiated tissue with no regeneration capacity. If damaged, myocardial cells will be replaced by fibrous connective tissue. OBJECTIVE: To rediscover the myocardial cells and to do a review for cardiac stem cells, in order to define the existence of myocardial cells. METHODS: A computer-based online research of CNKI and PubMed databases was performed to collect articles published between 2003 and 2014 with the key words of “cardiac stem cells, stem cells, cardiac regeneration” in Chinese and English, respectively. There were 82 articles after the initial survey, and finally 40 articles were included in result analysis. RESULTS AND CONCLUSION: Cardiac stem cells exist in the heart, and some surface markers of cardiac stem cells have been discovered. Cardiac stem cells for some diseases with myocardial cell injury have opened up a new way. Because of the small number of myocardial cells, how to purification, culture, identification and proliferation will be further studied to meet the need for regenerative medicine and tissue engineering. Cardiac stem cell research will open a new approach for cardiac tissue engineering. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics