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01 January 2012, Volume 16 Issue 1 Previous Issue    Next Issue

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Some problematic issues about mesenchymal stem cells and their clinical application Guo Zi-kuan 2012, 16 (1):  1-10.  doi: 10.3969/j.issn.1673-8225.2012.01.001 Abstract ( 415 )   PDF (1KB) ( 846 )   Save Till now, mesenchymal stem cells (MSCs) have been officially approved in a series of clinical trials to treat the patients with graft-versus-host disease, Crhon’s disease, liver fibrosis and osteoarticular injuries. However, before their wide use in clinical settings, the basic problematic issues about MSCs should be clarified. These key points, which will be here reviewed based on the recent advances on MSCs during the past several years, include the essences of MSCs inherently existing in vivo, the changes of their biological features after in vitro expansion, in vivo distribution after transplantation, the underlying mechanisms of MSCs therapy and its safety in clinical application. References | Related Articles | Metrics

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Construction and expression of pHSV1-TK- IRES2-EGFP eukaryotic vector in mouse bone marrow mesenchymal stem cells Wu Li-chuan1, Yang Kun1, Liu Yong-zhe1, Chen Peng1, Xu Wen-gui2 2012, 16 (1):  11-16.  doi: 10.3969/j.issn.1673-8225.2012.01.002 Abstract ( 203 )   PDF (508KB) ( 391 )   Save BACKGROUND: Up to date, thestudies regarding the construction of herpes simplex virus 1-thymidine kinase (HSV1-TK) and enhanced green fluorescent protein (EGFP) reporter gene co-expression vector and its expression in mouse bone marrow mesenchymal stem cells are few. OBJECTIVE: To construct a pHSV1-TK- IRES2-EGFP eukaryotic vector, and to observe the expression of HSV1-TK in mouse bone marrow mesenchymal stem cells.  METHODS: The HSV1-TK cDNA fragments were obtained by polymerase chain reaction (PCR) from pHSV106 and Bgl Ⅱ, Sal Ⅰ two restriction sites were added, after T vector cloning, the recombinant plasmid pHSV1-TK/18T was digested by two restrictive endonucleases, and then HSV1-TK cDNA was collected and recombined with eukaryotic vector pIRES2-EGFP by using gene recombination technique. The recombinant plasmid pHSV1-TK- IRES2-EGFP was transfected into mouse bone marrow mesenchymal stem cells in vitro with lipofectamine transfection reagent. RESULTS AND CONCLUSION: A length of 1 130 bp with Bgl Ⅱ and Sal Ⅰ target gene sequences was obtained by PCR. And the pHSV1-TK- IRES2-EGFP expression plasmid was constructed successfully. The TK/GFP green fluorescent protein was detected after transfection. At the same time, the expression of HSV1-TK mRNA was determined. The eukaryotic vector pHSV1-TK- IRES2-EGFP was successfully constructed, and effectively expressed HSV1-TK mRNA in mouse bone marrow mesenchymal stem cells in vitro. Related Articles | Metrics

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Efficacy of labeling rat bone marrow mesenchymal stem cells with 5-ethynyl-2’-deoxyuridine Shi Gui-xiu, Sun Li-hua, Zhang Yue-xin 2012, 16 (1):  17-21.  doi: 10.3969/j.issn.1673-8225.2012.01.003 Abstract ( 211 )   PDF (510KB) ( 619 )   Save BACKGROUND: A safe, efficient, and convenient labeling method with high sensitivity and strong specificity is needed to understand the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro and in vivo. OBJECTIVE: To investigate the efficacy of labeling BMSCs with 5-ethynyl-2’-deoxyuridin (EdU) and its effects on cell proliferation. METHODS: BMSCs were isolated by adherence method from bone marrow of rats and cultured to the third generation. BMSCs were seeded into 96-well plate with concentration of 5×107/L. BMSCs were labeled with EdU at different concentrations (0, 10, 20 and 50 μmol/L). The EdU-labeling positive rates and morphology of BMSCs in each group were detected at 24, 48 and 72 hours of culture, and the cell growth curves were obtained. RESULTS AND CONCLUSION: The EdU-labeling positive rates were gadually increased in each group as time went on (P < 0.05), and the differences in the increased EdU-labeling positive rates were not significant (P > 0.05). Good EdU-labeling positive rates could be obtained in the 10 μmol/L group at 72 hours of culture. The BMSCs growth curves of each group before and after labeling were “S”-shaped. This suggested that application of EdU labeling within the experimental concentrations for BMSCs in vitro is efficient and safe without effects on cell proliferation. Good labeling rate of BMSCs can be obtained through labeling with EdU in 10 μmol/L for 72 hours. Related Articles | Metrics

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Effect of mesenchymal stem cells conditioned medium on protection of hydrogen peroxide injured cardiomyocytes in rats Liu Xin-bin1, 2, Zhang Hong-chao2, Guo Zi-kuan3 2012, 16 (1):  22-26.  doi: 10.3969/j.issn.1673-8225.2012.01.004 Abstract ( 264 )   PDF (628KB) ( 358 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) transplantation can reduce myocardial infarct area and improve cardiac function. To date, most believe that MSCs play an important role in secreting biological factors. OBJECTIVE: To observe the influence of MSCs conditioned medium (MSCs-CM) on cardiomyocytes apoptosis induced by hydrogen peroxide (H2O2). METHODS: Rat MSCs and cardiomyocytes were isolated, cultured separately and identified. Preparation of MSCs-CM and H2O2 injured NRCs model. The rats were divided into four groups: control group, model group, MSCs-CM group and MSCs-CM+inhibitors of PI3K (Wortmannin) group. Lactate dehydrogenase (LDH) activity and apoptosis rates were detected by ELISA and flow cytometry separately in order to assess cardiomyocytes injury. RESULTS AND CONCLUSIONS: LDH activity in the model group was significantly higher than that in the other groups (P < 0.001); While the LDH activity in the MSCs-CM+Wortmannin group was also higher than that in the MSCs-CM group (P < 0.001). The apoptosis rate in the MSCs-CM group was lower than that in the MSCs-CM+ Wortmannin group (P < 0.01), and the apoptosis rate in the two former groups was lower than that in the model group (P < 0.01). It indicated that MSC-CM inhibited H2O2-induced apoptosis of NRCs partially via PI3K pathway. Related Articles | Metrics

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Isolation and charcaterization of human bone marrow mesenchymal stem cells using a whole bone marrow adherent culture technique Zhang Hao1, 2, Zhang Bin2, Cheng Mei1, Tao Yan-ling1, Hu Jiang-wei2, Xu Man2, Chen Hu2 2012, 16 (1):  27-30.  doi: 10.3969/j.issn.1673-8225.2012.01.005 Abstract ( 257 )   PDF (389KB) ( 265 )   Save Abstract BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are widely used in cell therapy and tissue regeneration, so their isolation and culture are a key technique for research and clinical application. OBJECTIVE: To isolate the BMSCs by whole bone marrow adherent culture technique and to establish an easy and effective method to isolate and culture BMSCs. METHODS: The BMSCs were isolated by whole bone marrow adherent culture technique, and purified and amplified by passage. BMSCs were induced to differentiate into adipocytes, osteocytes and chondrocytes in special differentiation condition.    RESULTS AND CONCLUTION: The purity of the BMSCs isolated by whole bone marrow adherent culture technique was more than 90%. The adherent cells displayed an abundant presence of CD73, CD90, CD105 and absence of Hematopoietic cell phenotype CD34, CD45 and HLA-DR which were detected by flow cytometry. The cell doubling time was (24.04±0.49) hours; Cell cycle showed that there was percentage of stem cells as G0 /G1 71.63% and S+G2+M 28.37% respectively. The induced and differentiated results showed than BMSCs could be differentiated into adipocytes, osteocytes and chondrocytes. Based on these findings, the whole bone marrow adherent culture technique is a better method to obtain MSCs from human bone marrow. Related Articles | Metrics

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Differentiation of human bone marrow derived mesenchymal stem cells into epidermal-like cells induced by human amniotic membrane Sa Ya-lian, Hua Ying-kun, Gao Jian-mei, Peng Zheng-guo, Dong Hong, Yan Xin-min 2012, 16 (1):  31-34.  doi: 10.3969/j.issn.1673-8225.2012.01.006 Abstract ( 202 )   PDF (416KB) ( 343 )   Save Abstract BACKGROUND: Cytokines are mostly used for inducing the differentiation of bone marrow mesenchymal stem cells (BMSCs) into epidermal-like cells, and human amniotic membrane is less applied to induce the differentiation of BMSCs through co-culture method. OBJECTIVE: To explore the feasibility of human BMSCs into epidermal-like cells by using human amnion membrane. METHODS: Human BMSCs were isolated, cultured, expanded and identified by cell morphology and expression of CD29, CD44 with flow cytometre. The 5th passage BMSCs were co-cultured with human amniotic membrane for 12 days. RESULTS AND CONCLUSION: Human BMSCs displayed a fibroblast-like morphology and the positive cells of CD29 and CD44 were 96.6% and 94.6%, respectively. After co-culture with human amnion for 12 days, the BMSCs differentiated into epidermal-like cells with diffuse Pan-CK, focal CK19. These findings indicate that human amnion can induce human BMSCs to differentiate into epidermal-like cells. Related Articles | Metrics

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Characteristics of fetal non-adherent bone marrow stromal cells  Xiao Long-yan1, Fu Jin-xiang1, Zhang Xue-guang2, Chen Qiu2, Zhang Hong1, Wang Pan-jun1, Sun Yu1, Wang Ming-yuan3, 2012, 16 (1):  35-38.  doi: 10.3969/j.issn.1673-8225.2012.01.007 Abstract ( 243 )   PDF (353KB) ( 335 )   Save Abstract BACKGROUND: There are non-adhesion stem cells in bone marrow stem cells, and the morphology and phenotype characteristics of the cells are similar to adhesion bone marrow mesenchymal stem cells.   OBJECTIVE: To explore the characteristics of fetal non-adherent bone marrow stromal cells (NA-BMSCs). METHODS: The fetal NA-BMSCs were cultivated by the method of adherent separation repeatly (“pull-offs”), the suspended cells were incubated in another culture bottle every 24 hours for 4 times. The growth curve of the passage 1, 3, 5 NA-BMSCs was detected by MTT assay, and the phenotype of the NA-BMSCs was determined with flow cytometry. Passage 1 NA-BMSCs were preformed with hematopoietic colony assay and passage 3 NA-BMSCs were preformed with osteogenic and adipogenic induction. RESULTS AND CONCLUSION: After four times’ “pull-off”, there were still asherent cells and colony formation; the resulting cells were continuing to culture for 7 days and those cells were the primary NA-MSCs (P0). The morphology of the cells was uneven; they highly expressed the integrin and the surface markers of endothelial cells with a middle level of CD106. Interestingly, they also expressed the surface markers of hematopoietic cells CD34. Those cells had multi-differentiation potency in vitro and they could differentiate into osteoblasts and hematopoietic cells under different inducing medium, but the capability of lipid differentiation was weak. Related Articles | Metrics

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Osteoblasts derived from human embryonic stem cells by CD271 magnetic beads in vitro Jiang Hong-hong1, Zhang Jin-li2, Li Fu-gui3, Wang Tao3 2012, 16 (1):  39-42.  doi: 10.3969/j.issn.1673-8225.2012.01.008 Abstract ( 229 )   PDF (377KB) ( 314 )   Save BACKGROUND: Studies on osteoblasts derived from human embryonic stem cells (hESCs) had made remarkable results, but the induced methods can produce a great number of effective osteoblasts derived from hESCs without other miscellaneous cells is not established. OBJECTIVE: To explore the feasibility of osteoblasts derived from hESCs in vitro by using CD271 magnetic beads sorting system. METHODS: CD271 positive cells were obtained from hESCs lines SYSU-2 cells through embryoid bodies formation at 6 days and attachment differentiation at 14 days by using CD271 magnetic beads sorting technology. Cell surface antigen expression, karyotype analysis and potential of differentiation were detected on CD271 positive cells. RESULTS AND CONCLUSION: CD271 positive cells exhibited regular spindle shape, which was similar to morphology of bone marrow-derived mesenchymal stem cells (BMSCs), and could be cultured in vitro about 14 generation remaining stable karyotype. At the same time, these cells expressed similar cell surface antigen marks (CD45 negative; CD73, CD105 and CD166 positive) with BMSCs. But differentiation potential identification showed that CD271 positive cells were solely induced osteoblasts formation. It has tremendous significance to study for the bone development mechanism and supply with enough, safe and efficient seed cells for bone tissue engineering. Related Articles | Metrics

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Guinea pig adipose mesenchymal stem cells differentiate into neuron-like cells Guo Wei1, Ji Bin1, Ma Yan2, Bi Xiao-juan2, Liu Li-zhong1 2012, 16 (1):  43-46.  doi: 10.3969/j.issn.1673-8225.2012.01.009 Abstract ( 154 )   PDF (376KB) ( 486 )   Save BACKGROUND: At present, most of the animal experiments about ear stem cells research are use the stem cells of the other animal species for the guinea pig model. The stem cells from guinea pig was rarely reported. OBJECTIVE: To explore the differentiated potential of the guinea pig adipose mesenchymal stem cells into neuron-like cells. METHODS: Guinea pigs adipose tissue were obtained, the adipose mesenchymal cells were isolated and cultured to the 3rd generation. (1) The absorbance values of cell proliferation from 1 to 10 days were tested by cck-8 kit. (2) The immunophenotype were detected by flow cytometry. (3) Differentiation of neuron-like cells was verified by nestin, glial fibrillary acidic protein (GFAP) and neurofilament protein 200 (NF200) immunofluorescence staining. RESULTS AND CONCLUSION: (1) The growth curve of adipose mesenchymal stem cells proliferation was similar to S-shaped, the first 3 days was the incubation period after passage, the cell populations entered logarithmic growth phase from the 4th day, after the 8th day they entered into the platform period. (2) The expression rate cells immune phenotype CD29 was 86.3%, CD44 was 55.5%, CD45 was 0.4%. (3) After differentiation of nerve cells, the morphology of cells was similar to neural cells, the results of nestin, GFAP and NF200 staining were positive. Guinea pig adipose mesenchymal stem cells are easy to isolation, culture and amplification in vitro. They have the potential to differentiate into neuron-like cells References | Related Articles | Metrics

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Immunogenicity and safety of mice adipose-derived stem cells Liang Shuang, Yuan Gui-feng, Zhong Yu-juan, Liu Jing, Chen Sen-zhou 2012, 16 (1):  47-50.  doi: 10.3969/j.issn.1673-8225.2012.01.010 Abstract ( 219 )   PDF (329KB) ( 413 )   Save BACKGROUND: Multiple sets of experiments confirmed that adipose-derived stem cells (ADSCs) can amplification in vitro, and can differentiated to three germinal layers under certain conditions and can be modified with gene. OBJECTIVE: To investigate the immunogenicity and safety of mice ADSCs after vein transplantation. METHODS: ADSCs were isolated from adipose tissue of mice; immunity markers of these cells were observed by flow cytometry. In vitro lymphocyte hyperplasy experiment was used to determine the effect of ADSCs on activated ability of lymphocyte. ADSCs were transplanted into mice model through caudal vein and observed the stimulatory function of ADSCs on T and B lymphocyte in vivo. Anatomy the mice model after transplantation and detect the tumor of the transplanted ADSCs. RESULTS AND CONCLUSION: ADSCs could culture in vitro and express the immunity markers of mesenchymal stem cells. When co-culture with lymphocyte, ADSCs could not stimulate the proliferation of lymphocyte efficiency. After transplantation through caudal vein, ADSCs could not stimulate immunological reaction of lymphocyte, but ADSCs could activate B lymphocyte and the humoral immunity. No tumor formation and tissue distortion were observed after transplantation. ADSCs have low immunogenicity, and could not stimulate the immune system of host cells and had a good safety. Related Articles | Metrics

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Isolation and culture of mouse adipose-derived stem cells and the expression of hematopoietic modulation factors before and after osteogenic induction Hao Dan1, Duan Xian-lin1, Bi Xiao-juan2, Chang Xiang-ping1, Ma Yan2, Qu Jian-hua1, Yuan Hai-long1, Jiang Ming1, 2 2012, 16 (1):  51-55.  doi: 10.3969/j.issn.1673-8225.2012.01.011 Abstract ( 298 )   PDF (478KB) ( 341 )   Save BACKGROUND: There are few reports about the isolation and culture of mouse adipose-derived stem cells in vitro and the expression of hematopoietic modulation factors before and after osteogenic induction. OBJECTIVE: To investigate the isolation and culture of mouse adipose-derived stem cells (ADSCs) and molecule level of hematopoietic modulation factors before and after osteogenic induction in vitro. METHODS: ADSCs were isolated from male C57BL/6 mouse by digesting adipose tissue with collagen typeⅠ, and expanded to three passages after primary culture in a control medium. Then the cells underwent the adipogenic and osteogenic induction. RESULTS AND CONCLUSION: ADSCs grew as spindle-shaped adherent cells when cultured in vitro, and could stably proliferate and be passed. The cells’ surface antigen phenotype was analyzed by flow cytometry, and the expression of CD29 was strongly positive, CD34 was lower, the expression of CD106 and CD49d was negative. ADSCs incubated in the adipogenic medium and osteogenic medium were stained positively. The expression of hematopoietic modulation factors-catenin was the highest, the stromal cell derived factor-1 and N-cadherin was followed by, the expression of the other factors such as stem cell factor, macrophage inflammatory protein-1α, ligand jagged1 and granulocyte macrophage colony-stimulating factor were all lower. It indicates that the cells from adipose tissue are ADSCs, and the expression of hematopoietic modulation factors is found before and after osteogenic induction, but has no significant difference.  Related Articles | Metrics

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Effects of human embryonic lung fibroblasts feeder layer transfected with leukemia inhibitory factor gene on the proliferation of umbilical cord blood CD34+ cells in vitro Xia Wei1, Yu Xin1, Jiang Jian1, Tian Li-na2, Chen Yao2, Miao Jing-cheng2 2012, 16 (1):  56-60.  doi: 10.3969/j.issn.1673-8225.2012.01.012 Abstract ( 260 )   PDF (334KB) ( 330 )   Save BACKGROUND: The number of hematopoietic cells in single copy cord blood is limited, and it is difficult to meet the needs of adults. How to effectively amplify cord blood hematopoietic stem/progenitor cells (HSPCs) is a hot research issue currently. OBJECTIVE: To construct gene-modified human embryonic lung fibroblast cells using human leukemia inhibitory factor (hLIF) and to observe the effect of transgenic cells on the amplification of HSPCs in vitro. METHODS: The feeder layer trancfected with recombinat adenouirus Ad-hLIF was established and objective gene was detected by using RT-PCR and ELISA. Umbilical cord blood CD34+ HSPC were separated using magnetic-activated cell sorting and the purity was detected using flow cytometry. Homing ability of amplified HSPC in vitro was detected by transmembrane migration assay. RESULTS AND CONCLUSION: Transgenic feeder cells were constructed successfully, and the expression of objective gene was verified by RT-PCR and ELISA. After culturing with feeder layer cells for seven days, the number of CD34+ HSPCs in the group containing hLIF was 15.73 much more than that in the group without hLIF. The expression rate of surface adhesion molecules was also still great. In vitro migration assay showed that the induced migration rate of hematopoietic cells in co-culturing with transgenic feeder layer was significantly higher than that of the control group, with a better homing ability. The feeder layer cells of adenovirus vector transfected with hLIF gene amplify umbilical cord blood CD34+ HSPC effectively in vitro and delay differentiation; what’s more, the CD34+ cells retains a great homing ability after amplifying in vitro. Related Articles | Metrics

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Stromal cell-derived factor-1/CXCR4 effects on ex vivo expanded bone marrow mesenchymal stem cells migration Wang Jin, Huang Liang-hu, Wang Qing-hua, Shi Xiao-hua, Dong Hui-yue, Tan Jian-ming 2012, 16 (1):  61-64.  doi: 10.3969/j.issn.1673-8225.2012.01.013 Abstract ( 321 )   PDF (375KB) ( 285 )   Save BACKGROUND: Exogenous human mesenchymal stem cells (MSCs) possess an ability to selectively migrate to the site of injury, and may further repair various damaged tissue. The mechanisms of directional migration remain unclear. OBJECTIVE: To examine the expression of the CXCR4 receptor in MSCs after long-term culture ex vivo, and to investigate the effects of stromal cell derived factor-1 (SDF-1) at various concentrations on MSCs migration. METHODS: MSCs were cultured ex vivo to passage 10, and then cell senescence was determined by β-galactosidase staining. The expression of CXCR4 receptor in MSCs was examined by RT-PCR and immunofluorescence staining. Transwell system was used to detect in vitro migration capacity of MSCs towards SDF-1. RESULTS AND CONCLUSION: Proliferation gradually decreased in all samples in the course of long-term culture ex vivo, and became restricted to senescent stages at passage 6 or 7. The expression of CXCR4 receptor was gradually lost with culture expansion. SDF-1 induced the migration of MSCs in a dose-dependent manner. Wang J, Huang LH, Wang QH, Shi XH, Dong HY, Tan JM. Stromal cell-derived factor-1/CXCR4 effects on ex vivo expanded bone marrow mesenchymal stem cells migration.Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(1): 61-64.     [http://www.crter.cn  http://en.zglckf.com] Related Articles | Metrics

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Effect of nerve growth factor and epidermal growth factor on proliferation of endogenous neural stem cells after traumatic brain injury Liu Yang1, Liu Wei-ping2, Wang Xiao-an2, Yang Yang2, Long Qian-fa2, Zheng Zhao-hui2, Zhang Yu-qi2, Zhong Jun2 2012, 16 (1):  65-69.  doi: 10.3969/j.issn.1673-8225.2012.01.014 Abstract ( 215 )   PDF (370KB) ( 433 )   Save BACKGROUND: Epidermal growth factor (EGF) and nerve growth factor (NGF) can promote the proliferation of neural stem cells, but their effects on endogenous stem cells after a traumatic brain injury in adult rats are studied little. OBJECTIVE: To investigate the effect of NGF and EGF on proliferation of endogenous neural stem cells after traumatic brain injury in rats. METHODS: Forty-eight adult SD rats were divided into three treatment groups and one control group at random, and the traumatic brain injury model was established with improved Feeney’s free-fall impacting method. Separate and combined injection of NGF and EGF were given to rats at 24 hours after traumatic brain injury. Normal saline was administrated into the rats in the control group. RESULTS AND CONCLUSION: The scores of forelimb placing test and beam balance test examines in the NGF, EGF or the combination group were superior to those of the control group, and the scores in the combination group were better than those in the NGF or EGF group (P < 0.05). Compared with the control group, BrdU-labelde cells were detected in the bilateral subependymal zones, the dentate gyrus of the hippocampus and damage region after traumatic brain injury in NGF, EGF or the combination treatment group increased more significantly; the expression in the combination group was better than in the NGF or EGF group (P < 0.05). Proliferation of endogenous neural stem cells and restoration of limbs function after traumatic brain injury can be promoted by NGF or EGF. Morever, this effect can be improved more significantly by the combination of NGF and EGF.   Related Articles | Metrics

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Human bone marrow mesenchymal stem cells transplantation induces neural regeneration and improves the recovery of nerve function in a rat stroke model Liu Tai-yun1, Xiong Fu2, Lin Jun1, Wang Chuan-Sen1, Zhang Cheng3 2012, 16 (1):  70-75.  doi: 10.3969/j.issn.1673-8225.2012.01.015 Abstract ( 228 )   PDF (409KB) ( 363 )   Save BACKGROUND: Studies have shown that allograft of bone marrow mesenchymal stem cells (BMSCs) or neural stem cells (NSCs) can improve the nerve function of rat stroke model.   OBJECTIVE: To investigate survival and migration of BMSCs in part of the infarction region of transplanted rat and the neural regeneration.  METHODS: Human BMSCs were transplanted into an immunosuppressed rat model of focal cerebral ischemia by vein. Behavioral recovery (neurological severity scores, NSS; rotarod test) and nerve function were also observed once a week. The rat were euthanized at 1-8 weeks after transplantation, and the expression of human nuclear antigen (HNA), marker of neural stem cells (Nestin) and nerve cell markers (Neuronal nuclei, cerebral tubulin, glial fibrillary acidic protein) were analyzed by immunohistochemistry staining, immunofluorescence staining and RT-PCR.  RESULTS AND CONCLUSION: At 2 weeks after BMSCs transplantation, the NSS score of rat stroke model was decreased significantly (P < 0.05), and the residence time of the rotarod test was significantly prolonged. At 1-8 weeks after BMSCs transplantation, HNA+ cells were identified in subependymal region, hippocampus dentate gyrus and the site around the infarction. At 1 and 2 weeks after BMSCs transplantation, Nestin positive cells and mRNA expression could be seen in subependymal region, hippocampus dentate gyrus (CA1, CA3) and the site around the infarction, and lasted for 8 weeks. Two weeks later, HNA, cerebral tubulin positive cells and mRNA expression could be seen in subependymal region, hippocampus dentate gyrus and the site around the infarction. At 4-8 weeks after BMSCs transplantation, a small amount of glial fibrillary acidic protein positive cells and mRNA expression could be seen in subependymal region, hippocampus dentate gyrus and the site around the infarction. The data indicate that human BMSCs can survive and migrate to focal cerebral ischemia of a rat stroke model by intravenous delivery, then differentiate into nerve cells, promote the formation of nerve cells and improve the recovery of neurological function. Related Articles | Metrics

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Angiogenesis in lower limb ischemia of diabetic rats after bone mesenchymal stem cells transplantation Wu Chun-peng1, Zhang Qian2, Fang Ning1, Cai Zheng3, Shi Rong-shu3, Liu Zu-lin1, Liu Jin-wei1, Zhang Tao1 2012, 16 (1):  76-80.  doi: 10.3969/j.issn.1673-8225.2012.01.016 Abstract ( 222 )   PDF (500KB) ( 383 )   Save BACKGROUND: Whether bone marrow mesenchymal stem cells (BMSCs) is the major source of seed cells to promote angiogenesis in lower limb ischemia. OBJECTIVE: To investigate the angiogenesis evidence in lower limb ischemia of diabetic rats after intramuscular injection of BMSCs. METHODS: A rat diabetic ischemic lower limb model was made by intraperitoneal injection of streptozotocin plus ligation of the bilateral femoral arteries. BrdU-labeled BMSCs were injected into 10 points of the left lower limb ischemic muscles such as rectus femoris and gastrocnemius beginning at ligation site. The same muscles of right lower limb were injected with equivalent normal saline as control. RESULTS AND CONCLUSION: Digital subtraction angiography imaging analysis showed that vasculature of bilateral legs in model rats could be observed at 2 weeks post-transplantation, and was obviously increased at 6 weeks after implantation of BMSCs, the vasculature in left leg was always higher than that in the right. At 2 weeks after transplantation, capillary densities of rectus femoris and gastrocnemius in left leg were higher than right leg (P < 0.05), and some implanted BMSCs with positive BrdU in vascular wall, which might have been incorporated into the vasculature. It is indicated that transplantation of BMSCs by local injection can improve blood supply and contribute to angiogenesis in lower limb ischemia of diabetic rats. Related Articles | Metrics

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Differentiation of neural stem cells into glial cells cultured in serum-free medium containing growth factors by using neurosphere assay Huang Fei, Zhao Ji-tong, Shen Qiang 2012, 16 (1):  81-84.  doi: 10.3969/j.issn.1673-8225.2012.01.017 Abstract ( 201 )   PDF (369KB) ( 301 )   Save BACKGROUND: Isolation and culture of neural stem cells in vitro and to master their biological characteristics are the basis of neural stem cells transplantation. OBJECTIVE: To isolate and culture cerebral cortex neural stem cells from rat embryos by neurosphere assay, and to observe the growth and differentiation characteristics of neural stem cells. METHODS: Neural stem cells were isolated from rat embryonic cerebral cortex and cultured in serum-free medium. Immunocytochemical staining was used to examine neural stem cells and differentiated cells markers. RESULTS AND CONCLUSION: Neural stem cells grew as free-floating neurospheres in the serum-free conditions, but spontaneous fusion occurred among some neurospheres. Immunocytochemical staining showed these neruospheres positively expressed nestin, and neural stem cells could differentiated into neurons, astrocytes and oligondendrocytes, but mainly differentiated into astrocytes. In serum-free medium containing growth factors, cerebral cortex neural stem cells could be isolated and cultured by using neurosphere assay. Related Articles | Metrics

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Effects of bone marrow mesenchymal stem cells transplantation in different ways on hippocanipal neurons following traumatic brain injury in rats Lü Jia-xi, Yin Hong 2012, 16 (1):  85-89.  doi: 10.3969/j.issn.1673-8225.2012.01.018 Abstract ( 153 )   PDF (441KB) ( 585 )   Save BACKGROUND: In recent years, application of bone marrow mesenchymal stem cells (BMSCs) transplantation in the central nervous system has become more widely, and the study of the best way to transplantation is important. OBJECTIVE: To investigate the effect of BMSCs transplantation in different ways on hippocanipal neuron following traumatic brain injury (TBI) in rats. METHODS: BMSCs were isolated from the bone marrow of 1-month-old SD rat. The BMSCs at passage 3 were labeled with PKH26 and were observed under fluorescence microscope. The percentage of the labeled cells was detected by flow cytometer. The model of hippocampal neuronal death following TBI was established by improved Feeney’s weight drop techniques. Sixty Wistar rats were divided into three groups randomly: Injured brain area transplantation group was injected with 10 μL normal saline contained BMSCs; Vena caudalis transplantation group was injected with 1 mL normal saline contained BMSCs through vein caudalis; TBI group was not treated. RESULTS AND CONCLUSION: Fluorescence microscope observation showed that the BMSCs labeled by PKH26 were spherical and presented with red fluorescence and uniform-distributed. Flow cytometric detection revealed that the percentage of labeled cells was over 99%. The learning and memory function of injured brain area transplantation group was significantly better than that of other two groups. The number of PKH26-positive cells and neurons after hematoxylin-eosin staining in injured brain area transplantation group was higher than that in vena caudalis transplantation group, and the number in vena caudalis transplantation group was higher that in TBI group (P < 0.05). RT-PCR approved that the expression of GAP-43 mRNA was highest in injured brain area transplantation group, a few in vena caudalis transplantation group and lowest in TBI group (P < 0.05). BMSCs transplantation plays an important role in protecting against hippocampal neuronal death following TBI. Injured brain area transplantation was significantly better than vena caudalis transplantation. Related Articles | Metrics

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Different quantities of neural stem cells transplantation in treating experimental cerebral infarction Wang Liang1, Cui Wei-yun2, Wang Xin-ping3, Wang Shi-min3, Wei Hui-jie2, Wang Dong2 2012, 16 (1):  90-94.  doi: 10.3969/j.issn.1673-8225.2012.01.019 Abstract ( 152 )   PDF (409KB) ( 273 )   Save BACKGROUND: Studies have confirmed that neural stem cells (NSCs) transplantation can promote nerve functional recovery in animals with cerebral infarction. OBJECTIVE: To observe the effect of different quantities of NSCs transplantation in rats with cerebral infarction and to find the minimum quantity of transplanted cells to promote nerve functional recovery effectively. METHODS: Embryonic brain tissues of Wistar rats were extracted and NSCs were cultured in vitro. The cerebral infarction models of Wistar rats were constructed by using suture method. At 7 days after cerebral infarction, cultured NSCs were extracted at 12 days. BrdU-labeled NSCs with 6×105, 8×105 and 10×105 were transplanted using stereotactic and micro-injection pump. A sham-transplanted group was established. Behavioral changes at 3 months after transplantation were evaluated by using prehensile traction test and inclined plane test. The differentiation of transplanted NSCs integrated with the surrounding host into neurons and glial cells was detected by using immunohistochemical staining in BrdU, neuron-specific enolase, glial fibrillary acidic protein. RESULTS AND CONCLUSION: NSCs could survive in the host brain after transplantation and differentiate into neurons and glial cells to promote functional recovery. At 3 months after NSCs transplantation, compared with the sham-transplanted group, the behavioral score was significantly increased in the transplanted group. The behavioral score was improved more significantly in the moderate quantity transplanted group than in the low quantity transplanted group, while there was no significant difference between the moderate quantity transplanted group and high quantity transplanted group. It is indicated that different quantities of NSCs transplantation in treatment of cerebral infarction can promote dysfunction recovery. The quantity of NSCs transplantation 8×105 can play better effect with less quantity transplantation. Related Articles | Metrics

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Transplantation of human amniotic epithelial cells in treating neonatal rats with hypoxic-ischemic brain encephalopathy Liu Hong-min, Chen Xu-dong, Hua Xin-yu 2012, 16 (1):  95-98.  doi: 10.3969/j.issn.1673-8225.2012.01.020 Abstract ( 163 )   PDF (384KB) ( 302 )   Save BACKGROUND: The previous studies showed that transplantation of human amniotic epithelial cells (hAECs) could improve neurological function, but the mechanism is unclear. OBJECTIVE: To explore the effects on neural stem cells and the expression of Neurogranin (Ng) of neonatal rats with hypoxic-ischemic encephalopathy, and the therapeutic effects of transplantation of human amniotic epithelial cells (hAECs). METHODS: Forty-six rats were randomly divided into sham operation control, model group, transplantation group. The 1×1012/L hAECs and PBS were respectively injected into the cerebral ventricle of the transplantation group and model group, and sham operation group did not treated. RESULTS AND CONCLUSION: Compared with model group, the neurological function was significantly improved in the transplantation group, NSE positive cells were detected in the transplantation group rat brains at 3 weeks following transplantation, Ng proteins expression in the model group was significantly lower than the sham operation group and the transplantation gorup at 3 weeks following transplantation. nestin proteins expression in the model group was higher than that in the sham operation group and lower than that in the transplantation group. Results indicated that hAECs injected into the cerebral ventricles could reduce the hypoxic-ischemic injury to neurons and improve neurological function. The mechanism was maybe achieved by autogenous neural stem cell proliferation, differentiation and synaptic regeneration stimulated by hAECs and its secretion. Related Articles | Metrics

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Human umbilical cord blood mononuclear cell transplantation for extensive anterior-wall acute myocardial infarction with cardiogenic shock and severe heart failure in one case★ Zhang Ming1, Yu Le2 2012, 16 (1):  99-102.  doi: 10.3969/j.issn.1673-8225.2012.01.021 Abstract ( 211 )   PDF (290KB) ( 279 )   Save BACKGROUND: Transplantation of the human umbilical cord blood mononuclear cells (HUCBCs) have received increasing attention, as a promising candidate for the cellular transplantation, but the majority of the existing studies are basic research. OBJECTIVE: To report a patient of extensive anterior-wall acute myocardial infarction with cardiogenic shock and severe heart failure, after treatment of HUCBCs transplantation. METHODS: A 73-year-old female patient with cardiogenic shock and severe heart failure after extensive anterior-wall acute myocardial infarction was treated with percutaneous coronary intervention (three scaffolds implantation) and medications, and she still appeared the symptoms of congestive heart failure, such as severe recurrent dyspnea. 2.4 × 108 HUCBCs (50 mL cell suspension) was injected into the infarcted myocardium through the left anterior descending artery by using coronary micro-guide catheter. RESULTS AND CONCLUSION: The patient reported profound clinical benefit including improvement of heart-failure-associated symptoms after the transplantation. Notably the patient did not experience the cell transplant-related side effects during 4 months of follow-up. The ejection fraction increased from 22% before the transplantation to 53% at 21 days after the transplantation. The B-type natriuretic peptide decreased from 1 730 ng/L before the transplantation, 854 ng/L after the transplantation to 264 ng/L at 21 days after the transplantation. The patient did not appear the symptoms of congestive heart failure, including dyspnea, chest distress and hypodynamia, she returned to daily activity at 4 months of follow-ups. Experimental findings indicate that the HUCBCs transplantation is an effective and safe means for patients cardiogenic shock and severe heart failure after acute myocardial infarction. Related Articles | Metrics

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In vitro directional differentiation of bone marrow mesenchymal stem cells into nerve cells induced by cistanche total glycosides Dong Ya-li, Li Jian-feng 2012, 16 (1):  103-106.  doi: 10.3969/j.issn.1673-8225.2012.01.022 Abstract ( 171 )   PDF (336KB) ( 861 )   Save BACKGROUND: Cistanche and its chemical composition play a neuroprotective role, and can promote recovery of neurological function. OBJECTIVE: To analyze the optimal dose of cistanche total glycosides for in vitro induction of bone marrow mesenchymal stem cells differentiating into nerve cells. METHODS: Bone marrow mesenchymal stem cells of Wistar rats were isolated and then cultured. β-mercaptoethanol and different concentrations of cistanche total glycosides were used to induce bone marrow mesenchymal stem cells. Bone marrow mesenchymal stem cells of Wistar rats were divided into blank control group, β-mercaptoethanol control group, cistanche total glycosides groups of 100, 200, 300 μg/L for observation. RESULTS AND CONCLUSION: After 7 days of induction, the number of living cells increased obviously in other groups, compared with the blank control group (P < 0.05); at the 3rd day of induction, Nestin, NSE-positive expression rate was significantly higher in cistanche total glycosides groups, compared with the blank control group (P < 0.05). 100, 200 μg/L cistanche total glycosides could effectively promote proliferation of bone marrow mesenchymal stem cells, and significantly promote the differentiation into nerve cells. 300 μg/L cistanche total glycosides could promote proliferation of bone marrow mesenchymal stem cells, but the effect was less than 100, 200 μg/L (P < 0.05). Related Articles | Metrics

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Effects of Chinese herbal compound on acute myeloid leukemia stem cells Liu Kui, Xu Rui-rong, Hu Shu-bo, Wang Xiao-ling, Cui Xing, Wang Yan, Wang Jing-yi 2012, 16 (1):  107-110.  doi: 10.3969/j.issn.1673-8225.2012.01.023 Abstract ( 243 )   PDF (330KB) ( 646 )   Save BACKGROUND: The existence of leukemic stem cells (LSCs) in leukemic patients is the basment how leukemia occurs and develops, drug resistance and recurrence happens. OBJECTIVE: To investigate the effects of Chinese herbal compound on the LSCs of acute myeloid leukemia (AML) patients. METHODS: CD34+CD38- LSCs were isolated from the bone marrow of 30 AML patients by immunomagnetic beads. The cells were divided into two groups: Control group were treated without medicine, while experiment group were treated by Chinese herbal compound preparation with terminal concentration of 100 mg/L. RESULTS AND CONCLUSION: The inhibition rate of Chinese herbal compound to LSCs was (36.660±0.018)%. Nuclear factor κB (NF-κB) expressed in 22 specimens, and the positive rate was 73.33% (22/30). The activity of NF-κB was tested by electrophoretic mobility. DNA-protein band grey values were analyzed by gel imaging system.The activity of NF-κB in the experiment group was decreased compared with that in the control group (P < 0.05) by comparison of the DNA-protein band grey values. The results showed that Chinese herbal compound may interfere in the growth of LSCs by inhibiting the activity of NF-κB.   Related Articles | Metrics

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Comparison of immunomodulatory effects between Stro-1+ and Stro-1- mesenchymal stem cells by micro-porous membrane Xu Wen1, Zhang Yi-zhuo1, Zhao Dan-dan2, Zhao Zhi-gang1, Yu Yong1, Zhao Wei-peng1 2012, 16 (1):  111-115.  doi: 10.3969/j.issn.1673-8225.2012.01.024 Abstract ( 287 )   PDF (329KB) ( 319 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) have been proved to posses the immunosuppressive effect on proliferation of lymphocytes. However, MSCs have a feature of nonuniformity. In addition, the immunosuppressive effect of Stro-1+ MSCs is obviously stronger than that of Stro-1- MSCs. OBJECTIVE: To investigate whether the significant difference in suppressive effect between Stro-1+ and Stro-1- MSCs still exists following the physical separation of MSCs from lymphocytes. METHODS: 1×104 and 3×104 Stro-1+ or Stro-1- MSCs were seeded in the lower chamber of transwell plate for 1-2 hours in order to achieve cell attachment, and then 1×104 reactive peripheral blood lymphocytes (PBLs) were incubated with stimulating PBLs of the same amount in the upper chamber of the transwell plate. After 5 days of culture, proliferation of the PBLs was measured by 3H-TDR incorporation. RESULTS AND CONCLUSION: The immunosuppressive effect of Stro-1+ MSCs was significantly stronger than that of Stro-1- MSCs when there was a direct connection between MSCs and PLBs. However, the immunosuppressive effects of the two subsets followed using transwell culture system were decreased than before. It is indicated that the direct connection between MSCs and PBLs may play a key role in the immunosuppressive effect of MSCs, and few soluble factors contribute a little during this. Furthermore, such “cross-talk” between cells is the prerequisite for MSCs to play the advantaged immune regulation.   Related Articles | Metrics

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Expression of N-methyl-D-aspartic acid receptor 2B in the subventricular zone of juvenile rats and its cell types  Zhang Wei1, Gao Jun-ying2, Xu Jing1, Cui Gui-yun3 2012, 16 (1):  116-120.  doi: 10.3969/j.issn.1673-8225.2012.01.025 Abstract ( 251 )   PDF (506KB) ( 336 )   Save BACKGROUND: N-methy-D-aspartic acid receptor (NR) 2B can express on neural stem cells. In-depth study of its expression in subventricular zone (SVZ) of juvenile rats can help to reveal the mechanisms of neural stem cell proliferation, differentiation, migration. OBJECTIVE: To detect the expression of NR2B in SVZ of juvenile rats and cellular localization of NR2B on the peak of expression. METHODS: The 7, 14, 21, 28 days postnatal SD rats were used to made 5 μm frozen sections of brain tissue. The expression of NR2B in SVZ and the cellular localization of NR2B at 14 days after birth were observed. RESULTS AND CONCLUSION: Immunohistochemistry showed that there were differential expressions of NR2B in SVZ of 7, 14, 21, 28 days postnatal rats and the expression peak of NR2B was on 14 day; The expression of NR2B in the lateral angle of lateral ventricle was higher than that of other parts; NR2B was co-localized with glial fibrillary acidic protein (GFAP), Nestin, β-Ⅲ tubulin, and ependymal cell was most obvious. However, there was no co-localization with NeuN in SVZ. The expression level of NR2B was changed following the different postnatal time. The results of double immunofluore staining implied that NR2B might play an important role in neurogenesis. Related Articles | Metrics

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A micro-stimulation method based on fish oocytes extracts induces mouse spleen cells to express stem cell mark antigen Ruan Guang-ping, Yao Xiang, Pang Rong-qing, Wang Jin-xiang, Ma Li-hua, Wang Qiang, Deng Yong-li, Pan Xing-hua 2012, 16 (1):  121-124.  doi: 10.3969/j.issn.1673-8225.2012.01.026 Abstract ( 231 )   PDF (271KB) ( 269 )   Save BACKGROUND: The mouse and human somatic cells can be induced by some special factors to a state of embryonic cells called as induced pluripotent stem cells. OBJECTIVE: To further enhance the efficiency of fish oocytes extracts on inducing somatic cells to differentiate into pluripotent stem cells. METHODS: We compared a new method (micro-stimulation, 0, 0.05, 0.1, 0.2, 0.4 g/L fish oocytes extracts) to improve the induction efficiency, and the traditional method (0, 0.1, 0.2, 0.4, 0.8, 1.2 g/L fish oocytes extracts) in cultured cells by adding fish oocytes extracts. RESULTS AND CONCLUSION: Compared with the traditional method, the new micro-stimulation method could induce spleen cells to express more stem cell marker antigens OCT-3/4, Nanog, SSEA-1. It is proved that the micro-stimulation method is a more efficient way to generate pluripotent stem cells from somatic cells. Related Articles | Metrics

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Effect of miniature pig platelet-rich plasma on osteogenic induction of periodontal ligament stem cells in vitro Zhang Yuan1, Zhong Liang-jun2, Zhang Peng-tao1, Zhang Yuan-ming3, Xu Yan4 2012, 16 (1):  125-129.  doi: 10.3969/j.issn.1673-8225.2012.01.027 Abstract ( 310 )   PDF (451KB) ( 299 )   Save BACKGROUND: Tissue engineering technology provides a new way for bone defects caused by periodontitis. OBJECTIVE: To explore the effect of platelet-rich plasma (PRP) on osteogenic induction of miniature pig periodontal ligament stem cells (PDLSCs). METHODS: Collected the Guizhou miniature pig’s venous blood and prepared the centrifuged PRP three times; tissue block method was used to culture the periodontal ligament cells. 0.8%, 1.0%, 1.2% concentration of PRP were co-cultured with PDLSCs for 3, 7, 14 and 21 days, respectively. No PRP addition as control group. RESULTS AND CONCLUSION: Alkaline phosphatase (AKP) activity reach a peak at the 14th day after induced with 0.8%, 1.0% and 1.2% PRP, and highest activity of AKP was induced by 1.0% PRP. AKP activity was reduced at the 21st day. Alizarin red staining showed the PDLSCs were positive at 21st day after osteogenic induction of PRP. Purified monoclonal cell growth curve entering the logarithmic growth phase since the 3rd day, the amount of cells reached a peak at the 8th day. PRP can induce osteogenic PDLSC performance, especially the 1.0% PRP induction showed the optimal efficiency in vitro.   Related Articles | Metrics

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Effects of interleukin-3 on the proliferation of c-kit+Lin-cells in bone marrow   Zhang Chun-xing1, Wang Fang1, Liao Cai-xian2, Wu Ya-qiong1, Meng Bing1 2012, 16 (1):  130-133.  doi: 10.3969/j.issn.1673-8225.2012.01.028 Abstract ( 334 )   PDF (370KB) ( 305 )   Save BACKGROUND: c-kit+Lin- bone marrow derived liver stem cells are similar to other hemopoietic stem cells and not rich in bone marrow, so it is difficult to meet the needs of scientific research. OBJECTIVE: To elucidate the effect of optimal cytokine combinations with interleukin-3 (IL-3) on the proliferation of c-kit+Lin- cells in bone marrow. METHODS: c-kit+Lin- cells were isolated from mouse bone marrow by using a high-gradient magnetic cell sorting system (MACS), and different concentrations of IL-3 were injected into the combinations of stem cell factor (SCF), hepatic growth factor (HGF), FLt-3 ligand (FL), leukemia inhibitor factor (LIF), thrombopoietin (TPO) and cultured for 7 days in a liquid culture system. The number of cells, multiples of c-kit+Lin- cell proliferation and the apoptosis rate were detected. RESULTS AND CONCLUSION: Each group could expand c-kit+Lin- cells rapidly, and the numbers of cells expanded were 7-19 times as large as the beginning’s number. But when the concentration of IL-3 increased and reached to a certain concentration, the c-kit+Lin- cells did not increased effectively. The expansion of total cells was 245.41 times in 40 μg/L IL-3 group, but the expansion of c-kit+Lin- cells was only 15.80 times. There was significant difference between the 40 μg/L IL-3 group and 20 μg/L IL-3 group (P < 0.05). And in the 40 μg/L IL-3 group, the apoptosis rate showed the lowest was 4.66%. IL-3 showed a stronger potential for expanding c-kit+Lin- cells. It showed synergistic actions with SCF, HGF, FL, LIF and TPO to expand c-kit+Lin- cells and the phenotype was not changed, but too much IL-3 could not increased the c-kit+Lin- cells effectively. 20 μg/L IL-3 was the best concentration to expand c-kit+Lin- cells. Related Articles | Metrics

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Construction and expression of mouse chemokine receptor-7 green fluorescent lentiviral expression vector in immature dendritic cells  Song Li-xiao1, Zhang Pu1, Li De-peng1, Zeng Ling-yu2, Chen Chong2, Pan Xiu-ying1, Xu Kai-lin1, Huang Yi-hong1 2012, 16 (1):  134-138.  doi: 10.3969/j.issn.1673-8225.2012.01.029 Abstract ( 296 )   PDF (363KB) ( 334 )   Save BACKGROUND: Chemokine receptor 7 (CCR7) plays a key role in launching and regulating the migration of dendritic cells (DCs) from peripheral tissueto the lymphatic system. But immature dendritic cells (imDCs)do not express CCR7. Therefore, imDCs carrying CCR7 possess great prospect for stronger immune tolerance.   OBJECTIVE: To construct green fluorescent protein (GFP) lentiviral expression vector with mouse CCR7 gene and to observe the expression of CCR7 gene in imDCs. METHODS: The CCR7 was amplified from the mouse thymus by RT-PCR and transfected into pCR-Blunt carrier. The CCR7 DNA fragment and IRES-GFP were cloned into lentiviral expression vector LV-Lac, to form recombinant lentivial plasmid LV-CCR7. Three plasmids of the lentivial system (recombinant lentivial plasmid LV-CCR7, package plasmid ΔNRF, envelope plasmid pVSVG) were co-transfected with package lentivirus by lipofectamine. The imDCs were infected by the recombinant lentivial, The morphology of transfected imDCs was examined under a fluorescent microscope and flow cytometry was used to determine the expression of CCR7. RESULTS AND CONCLUSION: The CCR7 fragment was amplified from cDNA of the mouse thymus and subcloned to pCR-Blunt carrier, and lentiviral expression vector LV-CCR7 was constructed successfully. After infected with 293 FT cells for 24 hours through three plasmid package system, positive expression of GFP was observed under fluorescence microscope. The titer of the lentivirus was above 108 U/L, and to obtain the recombinant lentivial with mouse CCR7 gene. The lentivial plasmid could infect imDCs effectively, a large amount of GFP and CCR7 expression was observed under fluorescence microscope and flow cytometry, respectively, and the positive rate was 50%. It indicates that GFP lentiviral vector LV-CCR7 with mouse CCR7 gene were successfully constructed and expressed in imDCs.   Related Articles | Metrics

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Effect of bone marrow msenchymal stem cells on the treatment of glioma    Cao Pei-chao, Wang Jian-jiao, Zheng Yong-ri 2012, 16 (1):  139-142.  doi: 10.3969/j.issn.1673-8225.2012.01.030 Abstract ( 270 )   PDF (383KB) ( 283 )   Save BACKGROUND: Bone marrow msenchymal stem cells (BMSCs) have become the ideal cell vector for the treatment of glioma. OBJECTIVE: To understand the research process and related mechanism of BMSCs in glioma therapy research. METHODS: We searched Pubmed database (1990/2011-01) with the key words “bone marrow mesenchymal stem cells, mesenchymal stem cells, Multipotent stem cells,Treatment of glioma,Gene therapy”. The basic, perspective and clinical research articles that related to the biological characteristics of the BMSCs and the effect of the BMSCs on the treatment of glioma was included. The duplicate articles were excluded. RESULTS AND CONCLUSION: More than 109 articles were screened out by computer according to inclusive and exclusive criteria, 34 documents of which were involved for analysis. Articles were described the biological characteristics of BMSCs, compared the animal experiments and the clinical research of BMSCs transplantation for treatment of glioma, and it confirmed that BMSCs as vectors for gene therapy of glioma had good development prospects in glioma treatment. Related Articles | Metrics

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Intravenous bone marrow mesenchymal stem cells homing pathways Wang Yang1, 2, Xiao Yang1 2012, 16 (1):  143-147.  doi: 10.3969/j.issn.1673-8225.2012.01.031 Abstract ( 181 )   PDF (408KB) ( 663 )   Save BACKGROUND: Mesenchymal stem cells (MSCs), as non-hematopoietic multipotent stem cells, are considered as the most promising stem cells for the treatment of a variety of diseases. But their low rate of homing to target tissues and organs limit the therapeutic efficacy when they are used in stem cell therapy. OBJECTIVE: To investigate intravenous bone marrow MSCs homing phenomena and mechanisms based on analysis of related literatures at home or abroad. METHODS: A computer-based online search of CNKI database, Wanfang database and PubMed database was performed with key words “Bone marrow mesenchymal stem cells, homing” in Chinese and in English, respectively. And the articles were mainly selected to be the most representative literatures during the last 10 years. Totally 33 literatures were included according to inspection and sorting. RESULTS AND CONCLUSION: Many studies showed that intravenous bone marrow MSCs interacted with the vessel wall and migrated across the endothelium, homing to many tissues and organs, especially damaged tissue, inflammation and tumor. This article was discussed from the intravenous MSCs homing phenomena and mechanisms. We describe the regulatory signaling molecules and receptors involved, such as chemotatic factor and growth factor, and adhesion molecules Related Articles | Metrics

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Stem cells treatment for osteoporosis: Possibility and feasibility Jia Yi-jie1, Tian Jing2 2012, 16 (1):  148-152.  doi: 10.3969/j.issn.1673-8225.2012.01.032 Abstract ( 278 )   PDF (430KB) ( 324 )   Save BACKGROUND: Treatment of osteoporosis with drug is not ideal. In recent years, more and more scientists try to use stem cells to treat osteoporosis. OBJECTIVE: To explore the treatment for stem cells on osteoporosis in order to promote its clinical application. METHODS: A computer-based online search of PubMed database and CNKI database between May 1997 and October 2011 was performed to search related articles with the key words of “stem cell, osteoporosis, bone metabolism, osteoporosisbone marrow mesenchymal stem cells (BMMSCs), Adipose-derived stem cells (ADSCs)” in English or in Chinese. Literatures related to treatment for stem cells on osteoporosis were selected; in the same field, the articles published lately in authoritative journals were preferred. RESULTS AND CONCLUSION: A total of 144 literatures were primarily selected, and 50 documents were involved for summary according to inclusion criteria. The treatment for stem cells on osteoporosis was mainly by increasing the number of osteoblasts. Commonly used methods were stem cells transplantation and induced the differentiation of stem cells. Stem cells treatment which has broad application prospects is an essential way for the treatment of osteoporosis, and it can avoid the side effects of drug therapy. Related Articles | Metrics

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Differentiation of adipose-derived stem cells into nerve cells and their transplantation for cerebral ischemia Zhu Yan-fei1, Wang Lin-lin1, Dong Wei-ren2 2012, 16 (1):  153-157.  doi: 10.3969/j.issn.1673-8225.2012.01.033 Abstract ( 263 )   PDF (423KB) ( 292 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs) from the mesoderm have the potential of multiple differentiations. It can differentiate to neurons under specific environments and then transplant into animal models of cerebral ischemia which can enhance the recovery of neurological function. OBJECTIVE: To make a review of ADSCs in the treatment of cerebral ischemia, and to analyze current problems and to prospect the future. METHODS: PubMed, CNKI, CAJ were retrieved by computer with keywords of “adipose-derived stem cells, adipose-tissue derived stem cells, adipose stem cells, differentiation, cerebral ischemia”. The language was limited for English and Chinese. The literatures included clinical research, basic research, and reviews. Repetitive researches were excluded, and 34 documents of them were involved for further analysis. RESULTS AND CONCLUSION: In recent years, many researches show that ADSCs can promote the neurological function after cerebral ischemia in different degree. But by now there is no report that ADSCs can be induced to mature neurons. How to differentiate to neurons accurately in vivo is a focus of ADSCs in the treatment of cerebral ischemia. Related Articles | Metrics

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Development of umbilical cord mesenchymal stem cells applied in bone tissue engineering Ha Cheng-zhi, Wang Da-wei 2012, 16 (1):  158-162.  doi: 10.3969/j.issn.1673-8225.2012.01.034 Abstract ( 217 )   PDF (635KB) ( 437 )   Save BACKGROUND: In bone tissue engineering, the studies about umbilical cord mesenchymal stem cells (UC-MSCs) as the new seed cells are gradually increased. OBJECTIVE: To review the isolation, culture, biology characteristics and the carrier used in bone tissue engineering of UC-MSCs. METHODS: A computer-based online search of literatures published between January 1999 to March 2011 related to the isolation, culture, biology characteristics and the carrier of UC-MSCs was performed in PubMed database and CNKI database using the key words of “umbilical cord stem cell, bone tissue engineering” in English and in Chinese. Finally, 46 articles were included according to inclusion criteria. RESULTS AND CONCLUSION: UC-MSCs possess multiple properties of embryon and adult stem cells. UC-MSCs have been proved to meet the criteria of MSCs published by the international society for cellular therapy. However, the isolation, culture, and differentiation of UC-MSCs are still unclear. There are many studies on single scaffolds as carrier, the research about composite carrier and injectable carrier may be the hotspot in the future. Related Articles | Metrics

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Stem cell therapy for nervous system diseases Wang Yu, Wang Ren-zhi 2012, 16 (1):  163-166.  doi: 10.3969/j.issn.1673-8225.2012.01.035 Abstract ( 362 )   PDF (326KB) ( 510 )   Save BACKGROUND: Stem cells have capacity of multipotential differentiation and self-renewal, which can stabilize internal environment under physical or pathophysical conditions. So stem cells have been considered as a useful tool in regenerative medicine. OBJECTIVE: To overview the advancement of the stem cells therapies on nervous system diseases. METHODS: A computer-based online search of PubMed databases was undertaken for the literature of stem cells therapies on nervous system diseases using the keywords of “stem cells, nervous system diseases, therapy”. Totally 298 articles were collected initially, and finally 54 articles were included in result analysis. RESULTS AND CONCLUSION: Stem cells are currently considered promising candidates for therapies on nervous system injuries and neurodegenerative diseases. Stem cells transplantation can replace the damaged cells and reconstruct neural network, which can improve neural function. Wang Y, Wang RZ. Stem cell therapy for nervous system diseases.Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(1): 163-166.     [http://www.crter.cn  http://en.zglckf.com]   Related Articles | Metrics

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Genetic engineering and cell engineering for repair of damaged atrioventricular conduction or construction of artificial atrioventricular bypass Dong Hao1, Li Ping1, Li Xuan2 2012, 16 (1):  167-170.  doi: 10.3969/j.issn.1673-8225.2012.01.036 Abstract ( 365 )   PDF (396KB) ( 324 )   Save BACKGROUND: Using cell-engineering and gene technology to treat bradyarrhythmia has features such as not requiring external energy resource and not being regulated by the neural hormone. It is a way which is more similar to the nature human body. These advantages are incomparable to drugs and electronic pacing. OBJECTIVE: To review the development and existing problems in using genetic engineering and cell engineering to treat bradyarrhythmia. METHODS: A computer online search was performed to retrieve papers published between 2000-01 and 2011-06 in PubMed and CNIK database. RESULTS AND CONCLUSION: Genetic engineering healing bradyarrhythmia paid close attention to β2-adrenergic receptor and Kir2.1 first. And HCN, AC-VI and Tbx3 are the most popular ones in recent research. The stem cell transplantations including the stem cell modified by gene, the combination of cells, iPS and CSCs have displayed their great potential in the treatment of bradyarrhythmia. The obvious alternative here would be to build an atrioventricular bypass tract to permit the normal sinus nodal impulse to propagate to the ventricle. However, there are still a lot of key issues to deal with, such as the optimization of the atrioventricular delay, the program design of automatically responding and the position of the artificial bypass tract.   Related Articles | Metrics

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Effect of umbilical cord blood mesenchymal stem cells transplantation on immune function of spinal cord injury patients Zhao Ning1, Yang Wan-zhang1, Zhang Min1, Sheng You-xiang1, Tang Ying2 2012, 16 (1):  171-174.  doi: 10.3969/j.issn.1673-8225.2012.01.037 Abstract ( 279 )   PDF (358KB) ( 373 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) transplantation has certain immunogenicity. However, there was no in-depth and system reports about the effect of MSCs transplantation on spinal cord injury patient’s immune function. OBJECTIVE: To observe the effects of umbilical cord blood MSCs (UCB-MSCs) transplantation on immune function of spinal cord injury (SCI) patients. METHODS: Sixty-one spinal cord injury (SCI) patients were treated by intervenous drop infusion and lumbar subarachnoid space puncture to UCB-MSCs. Flow-cytometry and immunoturbidimetry (ITM) were used to detect the content of T lymphocytic subgroup and immune globulin and the alexinic in plasma of all the patients prior and post-transplantation. RESULTS AND CONCLUSION: Compare with pre-transplantation, the content of CD3+, IgA and IgG was decreased and the differences were significant; The content of CD4+, CD8+, CD4+/CD8+, IgM, C3 and C4 was also changed and the differences were not significant. It indicates that UCB-MSCs transplantation could not activate the immunological response of cell and humoral immunity, so the transplantation is safety. Negative accommodation is exist, but the relevant indicators need a further improvement, expand samples, and clearly relevant mechanism.  Related Articles | Metrics

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Umbilical cord blood stem cell transplantation for the treatment of diabetic foot in 23 cases Zhu Lü-yun, Wang Guang-yu, Ma Li-cheng, Hu Li-ye, Li Xiao-ling, Yang Shao-ling, Shan Wei, Hou Rong-rong 2012, 16 (1):  175-178.  doi: 10.3969/j.issn.1673-8225.2012.01.038 Abstract ( 273 )   PDF (264KB) ( 326 )   Save BACKGROUND: Transplantation of autologous bone-marrow mononuclear cells or umbilical cord derived mesenchymal stem cells for treatment of diabetic foot has been widely developed at present, which showed good curative effect on ischemic lesions and peripheral neuropathy. OBJECTIVE: To observe the effect of umbilical cord derived mononuclear cells on diabetic ischemic lesions and peripheral neuropathy. METHODS: Twenty-three patients with diabetic foot received umbilical cord derived mononuclear cells injection into both lower extremities intramuscular (1-3 mL/site) for 3 cm distance after dilution. The number of implanted cells was 1.23×108-1.06×109 per leg (totally 38 limbs). RESULTS AND CONCLUSION: There was no significant difference in ankle/brachial index after treatment. There were significant differences in angina cruris, skin temperature and transcutaneous oxygen partial pressure after 3 or 6 months treatment. After treatment for 6 months, the pain score and thermoesthesia score were improved and there were significant differences in rational symptom score, clinical examination score, the value of pallesthetic sensibility and the conduction velocity of motor/sensory nerves. The clinical symptoms and objective indexes of diabetic ischemic lesions and peripheral neuropathy were improved after umbilical cord derived mononuclear cells implantation. Related Articles | Metrics

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Levels of peripheral blood stem cell factors in patients with chronic kidney diseases Qiu Fang-xin, Zhang Wei, Liang Yao-xian, Liu Xue-mei 2012, 16 (1):  179-182.  doi: 10.3969/j.issn.1673-8225.2012.01.039 Abstract ( 268 )   PDF (305KB) ( 287 )   Save BACKGROUND: Stem cell factor (SCF) is a multifunctional cytokine, which plays a crucial role in the process of renal interstitial fibrosis. OBJECTIVE: To investigate the changes and clinical significance of serum SCF in patients with chronic kidney disease (CKD). METHODS: Serum levels of SCF in 22 healthy controls and 116 patients with CKD were detected by enzyme-linked immunosorbent assay. The correlation between serum levels of SCF and renal damage degree, anemia, lipid metabolism disorders, cardiovascular disease, uremic pruritusda was analyzed. RESULTS AND CONCLUSION: Serum levels of SCF in different CKD groups were significantly higher than that in control group (P < 0.01). Serum SCFs increased with the progression of renal dysfunction (P < 0.05), which were positively related to staging of CKD, serum creatinine, blood urea nitrogen, blood high C-reactive protein, lipids and parathormone, while negatively related to hemoglobin. The results suggested that serum SCF may play an importaut role in progression and medical camplications of CKD. Related Articles | Metrics

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Pathogenesis of chronic aplastic anemia suffering from kidney yin deficiency through the maternal genetics Cui Xing1, Zhang Wen-jing2, Cai Zhi-guo3, Xu Rui-rong1, Liu Fei1, Wang Jing-yi1, Liu Kui1 2012, 16 (1):  183-187.  doi: 10.3969/j.issn.1673-8225.2012.01.040 Abstract ( 269 )   PDF (666KB) ( 298 )   Save BACKGROUND: Several laboratories have reported unexpectedly large number of mitochondrial mutations in leukemia. But the direct relationship between mitochondrial mutations and chronic aplastic anemia (CAA) has not studied yet. OBJECTIVE: To study the mitochondrial mutations of CAA suffered from kidney yin deficiency and kidney yang deficiency, and to investigate the nature of maternal genetic: the relationship between mitochondrial and the occurrence and development of CAA suffered from kidney yin deficiency in order to further study the pathogenesis of CAA. METHODS: The bone marrow and the oral epithelium were obtained from 10 patients with CAA suffered from kidney yin deficiency and 5 patients with CAA suffered from kidney yang deficiency. DNA was extracted and underwent the entire sequencing of the mitochondrial DNA and compared the mitochondrial genome. RESULTS AND CONCLUSION: The entire sequencing of mitochondrial DNA in CAA suffer from kidney yin deficiency showed that the mutations were occurrence in the areas that closely related with mitochondrial oxidative respiratory chain, it included the reduced nicotinamide adenine dinucleotide dehydrogenase 1-2 and 4-6 and cytochrome B gene. However, the mitochondrial mutations in CAA suffered from kidney yang deficiency were not obvious. We are led to conclude that mitochondrial gene mutation can change the expression of respiratory chain enzyme complex in CAA patients, which results in energy metabolism impairment may participate in the physiological and pathology processes of hematopoietic failure. Functional impairment of mitochondrial respiration chain induced by gene mutation may be an important reason of hematopoietic failure in CAA. And this change is closely related to maternal inheritance and kidney yin deficiency. Related Articles | Metrics