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05 November 2013, Volume 17 Issue 45 Previous Issue    Next Issue

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In vitro neural-like cell differentiation from bone marrow mesenchymal stem cells: Co-culture versus chemical induction Xu Li-li, Kuang Tao, Zhang Jia 2013, 17 (45):  7821-7821.  doi: 10.3969/j.issn.2095-4344.2013.45.001 Abstract ( 118 )   PDF (2561KB) ( 479 )   Save BACKGROUND: A variety of methods have been used to induce bone marrow mesenchymal stem cells to differentiate into nerve cells. However, the percent of differentiated nerve cells is different using varied induction methods for different time periods. Appropriate induction condition is essential for induced differentiation of mesenchymal stem cells. OBJECTIVE: To find a suitable and practical method of inducing bone marrow mesenchymal stem cells into nerve cells in vitro by comparing chemical induction and co-culture methods. METHODS: Rat bone marrow mesenchymal stem cells were isolated by density gradient method and cultured by chemical induction and co-culture methods, respectively. Cell number, cell morphology, and specific cell surface markers were compared. RESULTS AND CONCLUSION: A large number of adherent cells with processes and radial growth, positive for neuron specific enolase staining were observed in both groups. Cells with typical nerve cell structure and a large number of processes were observed at 5 days in co-culture group, and the positive rate for neuron specific enolase staining was (50.82±2.46)%. Neural-like cells with processes were found at 7 days in chemical induction group, and the positive rate for neuron specific enolase staining was (43.56±1.74)%. Results showed that bone marrow mesenchymal stem cells were inducted into neural-like cells with more processes, which formed connection earlier cultured in co-culture compared with chemical induction. Moreover, the positive rate for neuron specific enolase staining was higher in co-culture group compared with chemical induction group. Figures and Tables | References | Related Articles | Metrics

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Establishment of human-rhesus chimeric liver using adult bone marrow mesenchymal stem cells He Bao-li, Ma Li-hua, Chen Li-ling, Liu Ru-wen, Yang Ren-hua 2013, 17 (45):  7827-7833.  doi: 10.3969/j.issn.2095-4344.2013.45.002 Abstract ( 90 )   PDF (2728KB) ( 327 )   Save BACKGROUND: Human-mammal chimeric liver chimera has been a vital significance for the proliferation and differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To establish an animal model of human-rhesus chimeric liver using adult bone marrow mesenchymal stem cells. METHODS: Adult bone marrow mesenchymal stem cells were isolated, purified and cultured for the sixth generation. The number of bone marrow mesenchymal stem cells was no less than 5×108. Bone marrow mesenchymal stem cells labeled with green fluorescent protein were transplanted into the liver of the embryo rhesus with pregnancy of 10 weeks under guided by type-B ultrasound. At the 1st and 3rd months of birth, the liver tissue of the infant rhesus was taken for biopsy. After routine pathological section, histological specimens were observed under fluorescence microscope to confirm if there were adult bone marrow mesenchymal stem cells positive for green fluorescent protein and their distribution, and detected by immunohistochemical staining to identify if human albumin expressed in the liver of infant rhesus. RESULTS AND CONCLUSION: Fluorescence microscope observation indicated that at the 1st and 3rd months after birth, there were surviving bone marrow mesenchymal stem cells derived from human with green fluorescence in the liver of infant rhesus, and these cells migrated to form more concentrated distribution. The immunohistochemical results demonstrated that functional liver cells expressing human albumin were observed in the liver of infant rhesus at the 1st and 3rd months after birth, and their distribution was in accordance with bone marrow mesenchymal stem cells with green fluorescence. Human-rhesus chimeric liver can be established using adult bone marrow mesenchymal stem cells, which can generate functional liver cells in the liver of infant rhesus. Figures and Tables | References | Related Articles | Metrics

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Differentiation of bone marrow mesenchymal stem cells into nucleus pulposus-like cells after co-culture with nucleus pulposus cells Wei Ge-jin, Huang Yu-qiang, Qin Wan-an, Liao Chang-chuan, Lin Zhou-dan 2013, 17 (45):  7834-7839.  Abstract ( 105 )   PDF (3146KB) ( 373 )   Save BACKGROUND: Bone marrow mesenchymal stem cells in recent years have been widely used as seed cells for nucleus pulposus tissue engineering. Appropriate extracellular microenvironment and growth factors are required for the induction of bone marrow mesenchymal stem cells differentiating into nucleus pulposus cells. OBJECTIVE: To observe the differentiation of rabbit bone marrow mesenchymal stem cells into nucleus pulposus-like cells in vitro induced by biological inducers and co-culture conditions. METHODS: Rabbit bone marrow mesenchymal stem cells and nucleus pulposus cells were isolated and cultured using monolayer culture method, respectively. Passage 3 bone marrow mesenchymal stem cells were placed in the Transwell culture plates for co-culture with the passage 3 nucleus pulposus cells and then were induced biologically to differentiate into the nucleus pulposus-like cells for 21 days. Bone marrow mesenchymal stem cells were cultured alone in the high-glucose Dulbecco’s modified Eagle’s medium as negative controls. RESULTS AND CONCLUSION: The expressions of CD29 and CD44 were positive and CD34 and CD45 were negative in the bone marrow mesenchymal stem cells. After 21 days of co-culture with nucleus pulposus cells, the morphology of bone marrow mesenchymal stem cells changed obviously from polygonal to oval shape, and collagen type Ⅱ granules were increased significantly. Reverse transcription-PCR results showed that expression of collagen type Ⅱ mRNA was significantly increased after induction. These findings indicate that bone marrow mesenchymal stem cells can be induced into differentiate into nucleus pulposus-like cells in co-culture plus biological inducer conditions in vitro. Figures and Tables | References | Related Articles | Metrics

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Osteogenic differentiation of human synovium-derived mesenchymal stem cells from osteoarthritic knee joints Rui Yun-feng, Lin Yu-cheng, Chen Hui, Wang Chen 2013, 17 (45):  7840-7846.  doi: 10.3969/j.issn.2095-4344.2013.45.004 Abstract ( 107 )   PDF (3056KB) ( 720 )   Save BACKGROUND: Mesenchymal stem cells are commonly used in tissue engineering, while whether synovium-derived mesenchymal stem cells from human knee joints can make a role in repair and regeneration of bone tissue as the appropriate seed cells need to be further verified. OBJECTIVE: To study the osteogenic differentiation potential of synovium-derived mesenchymal stem cells which were harvested from human knee joint with end-stage osteoarthritis in vitro. Meanwhile, to identify the osteogenic characteristics of these induced synovium-derived mesenchymal stem cells. METHODS: Cell populations were enzymatically released from the synovial membrane obtained from total knee arthroplasty. Nucleated cells were plated at an appropriate density (200 cells/cm2) for expansion at the maximum rate without colony-to-colony contact. Monoclone was obtained by selecting as primary synovium-derived mesenchymal stem cells. After primary cultured in control medium and expanded to three passages, synovium- derived mesenchymal stem cells were subjected to in vitro assays to investigate their osteogenesis potential in osteogenic medium containing dexamethasone, β-glycerophosphate and ascorbic acid. RESULTS AND CONCLUSION: Nucleated cells from the synovial membrane formed single cell-derived colonies, which were of polygon shape and star shape, uniform in size. After three passages, homogeneous populations of fibroblast-like cells were observed. Under appropriate culture conditions, synovium-derived mesenchymal stem cells were induced to differentiate to the osteocyte lineages which had typical “slabstone” appearance of osteoblasts. Osteogenesis was stained positively for alkaline phosphatase staining at day 7 and formed mineralized nodular structures at day 21, which was confirmed by Alizarin red staining. Alkaline phosphatase activity assay showed a rise after the osteogenesis induction and reached the peak at day 7. Expressions of osteocyte specific genes, such as collagen type Ⅰ, Runx2, bone-binding protein and osteopontin, were all detected. These genes were expressed positively in osteogenic medium, and the mRNA expressions of collagen type Ⅰ, Runx2, bone-binding protein and osteopontin were enhanced significantly after 21 days. Our study demonstrates that synovium-derived mesenchymal stem cells isolated from knee joint of end-stage osteoarthritis patients could be induced into osteoblasts in vitro, and these induced cells have typical osteogenesis characteristics. Synovium-derived mesenchymal stem cells may play a role in the regenerative response during the process of bone injury, which are promising candidates for bone tissue engineering. Figures and Tables | References | Related Articles | Metrics

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Stably upregulating expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells Zhou Lian-zhong, Cui Cheng-hua, Feng Ya, Xing Shuang-chun, Zhai Li-jie 2013, 17 (45):  7847-7854.  doi: 10.3969/j.issn.2095-4344.2013.45.005 Abstract ( 99 )   PDF (3385KB) ( 342 )   Save BACKGROUND: Chondromodulin-Ⅰ is expressed mainly in the cartilage, but it is little expressed in mesenchymal stem cells. Combined with the previous study of our group, we concluded that chondromodulin-Ⅰ maybe play an important role in inducing mesenchymal stem cells into chondrocytes accurately. OBJECTIVE: To construct an expression plasmid stably carrying chondromodulin-Ⅰ to up-regulate the expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells. METHODS: Specific primers were designed in rat cartilage for chondromodulin-Ⅰ gene, then the pcDNA3.1 (+) plasmid expression vector was digested by enzyme and directional connected gene to construct pcDNA3.1(+)/ChM-Ⅰ expression vector. Bone marrow mesenchymal stem cells were obtained from rats using the method of density gradient centrifugation combined with adherent culture. Recombinant plasmid pcDNA3.1(+)/ChM-Ⅰ was transfected into rat bone marrow mesenchymal stem cells with liposome method, and G418 selection was used for stable screen of transfected cells. Reverse transcription-PCR and western blot were used to detect chondromodulin-Ⅰ expression in cell lines. RESULTS AND CONCLUSION: The positive clones were digested by enzyme and were identified and sequenced. The results showed that the reality length and sequence of chondromodulin-Ⅰ gene were consistent with the theoretical values, and reading frame was correct. Reverse transcription-PCR and western blot results showed that the expressions of chondromodulin-ⅠmRNA and protein were markedly up-regulated in bone marrow mesenchymal stem cells. Recombinant plasmid pcDNA3.1(+)/ChM-I was successfully constructed, and transfected into rat bone marrow mesenchymal stem cells. After G418 selection, expression of chondromodulin-Ⅰ was up-regulated stably in rat bone marrow mesenchymal stem cells. Figures and Tables | References | Related Articles | Metrics

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Comparison of CM-DIL and DAPI labeled bone marrow-derived mesenchymal stem cells Shang Qing-qing, Li Kai, Zhou Jian-ye, Hu Sheng-shou 2013, 17 (45):  7855-7860.  doi: 10.3969/j.issn.2095-4344.2013.45.006 Abstract ( 239 )   PDF (1887KB) ( 657 )   Save BACKGROUND: Cell marker technology has been widely applied in many studies concerning cell transplantation. Chlormethylbenzamido-1,1-dioctadecyl-3,3,3’3’-tetramethylin-docarbocyamine (CM-DIL) and 4’,6-diamidino-2-phenylindole (DAPI) are commonly used for labeling cells. To our knowledge, there are few reports on comparing the two fluorescent dyes. OBJECTIVE: To compare the effects of CM-DIL and DAPI on labeling bone marrow mesenchymal stem cells in vitro and in vivo. METHODS: Isolation and expansion of bone marrow-derived mesenchymal stem cells were performed according to attachment culture. The cells were labeled by CM-DIL and DAPI, respectively. Cell viability was assessed via trypan blue exclusion assay. Growth curves of bone marrow-derived mesenchymal stem cells were depicted using MTS assay. The reduction of fluorescent intensity was observed under an inverted fluorescent contrast phase microscope from passage 1 to passage 3 after labeling. Myocardial infarction was induced by left anterior artery ligation in Sprague-Dawley rats. One week later, bone marrow-derived mesenchymal stem cells labeled by CM-DIL or DAPI were injected randomly into the border area of infarct myocardium. After 3 days, transplanted cell distribution was examined under the fluorescent microscope through paraffin sections and frozen sections respectively. RESULTS AND CONCLUSION: In vitro, bone marrow-derived mesenchymal stem cells labeled by both CM-DIL and DAPI showed decreased cell proliferation during the early period; the percentage of fluorescent-positive cells was approximately 100% in the two groups; however, the fluorescent intensity was significantly reduced from passage 1 to passage 3 in bone marrow-derived mesenchymal stem cells labeled by DAPI. In vivo, the transplanted cells were detected in a concentrated way both on the paraffin sections and frozen ones; the background color of frozen sections was lower in the CM-DIL group than in the DAPI group; false positive results of fluorescent expression could be eliminated in the CM-DIL group by using fluorescent mounting medium with the fluorescence of DNA staining. These data indicates that CM-DIL is more appropriate to in vivo tracing cells than DAPI. Figures and Tables | Related Articles | Metrics

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Influence of different oxygen partial pressures on cytokines secreted from human adipose-derived stem cells Jiang Yi-yao, Liu Xiao-cheng, Pei Yu, Zhu De-lin 2013, 17 (45):  7861-7868.  doi: 10.3969/j.issn.2095-4344.2013.45.007 Abstract ( 146 )   PDF (1760KB) ( 682 )   Save BACKGROUND: Effects of different oxygen partial pressures on cytokine secretion of human adipose-derived stem cells have been differently reported. These differences may be caused by varying oxygen partial pressures.  OBJECTIVE: To investigate the influence of different oxygen partial pressures on cytokines secreted from human adipose-derived stem cells.  METHODS: Human adipose-derived stem cells were cultured in vitro and identified by its immunophenotype. Human adipose-derived stem cells were divided into five groups and cultured under different oxygen partial pressure conditions (1%, 3%, 5%, 10%, 21%) for 24 hours, respectively. With quantitative real-time PCR and enzyme linked immunosorbent assay, the secretion of cytokines, vascular endothelial growth factor, hepatocyte growth factor, nerve growth factor, keratinocyte growth factor, from human adipose-derived stem cells were analyzed on the gene and protein levels. RESULTS AND CONCLUSION: Human adipose-derived stem cells were positive for CD71, CD73, CD90, CD105 and negative for CD34, CD45, CD54, HLA-DR. From the aspect of gene level, hypoxia (1%, 3% O2) promoted the expression of vascular endothelial growth factor and nerve growth factor from human adipose-derived stem cells (P < 0.01), and significantly elevated the expression of hepatocyte growth factor (P < 0.05); however, there was no significant influence on keratinocyte growth factor under hypoxia (P > 0.05). Based on the protein level, protein secretion of hepatocyte growth factor and vascular endothelial growth factor from human adipose-derived stem cells was increased under hypoxia (P < 0.01), but no changes occurred in nerve growth factor and keratinocyte growth factor. After cultured under hypoxic environment, human adipose-derived stem cells were promoted to express gene vascular endothelial growth factor, hepatocyte growth factor and nerve growth factor, as well as to secrete protein keratinocyte growth factor and vascular endothelial growth factor. Figures and Tables | References | Related Articles | Metrics

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Umbilical cord mesenchymal stem cells undergoing hypoxic preconditioning can differentiate into vascular endothelial-like cells Hao Xiao-juan, Zhu Lü-yun 2013, 17 (45):  7869-7876.  doi: 10.3969/j.issn.2095-4344.2013.45.008 Abstract ( 89 )   PDF (3103KB) ( 365 )   Save BACKGROUND: Incidence of diabetic lower extremity vascular disease is increasing, so how to improve blood vessels of the lower limbs and increase angiogenesis becomes the research focus. Umbilical cord mesenchymal stem cells have been employed clinically via local intramuscular injection, but the specific therapeutic effect and mechanism are not clear. OBJECTIVE: To investigate the effects of hypoxic preconditioning and cobalt chloride medium on the differentiation of umbilical cord mesenchymal stem cells into vascular endothelial-like cells in vitro. METHODS: Umbilical cord mesenchymal stem cells were isolated and cultured, and then treated with different concentrations of cobalt chloride. Enzyme linked immunosorbent assay was used to detect levels of basic fibroblast growth factor and vascular endothelial growth factor gene in cell supernatants, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to detect cell proliferation. The safety of umbilical cord mesenchymal stem cells before and after cobalt chloride induction was detected using chromosome. The umbilical cord mesenchymal stem cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10 μg/L vascular endothelial growth factor, 10 μg/L basic fibroblast growth factor and 10% fetal bovine serum, which were induced to differentiate into vascular endothelial-like cells. Endothelial-like cell phenotype CD31 and von Willebrand factor were identified before and after induction, through the observation of three-dimensional model of angiogenesis, the angiogenic capacity of umbilical cord mesenchymal stem cells was determined. RESULTS AND CONCLUSION: Umbilical cord mesenchymal stem cells strongly expressed the surface marks. After the cobalt chloride induction, the proliferation of umbilical cord mesenchymal stem cells was positivelycorrelated with the time of induction. Based on the levels of vascular endothelial growth factor and basic fibroblast growth factor, the optimal concentration of cobalt chloride was 200 µmol/L. Chromosome detection showed the stability of cells after cobalt chloride induction. After induction, CD31 and von Willebrand factor were strongly expressed. Three-dimensional observation showed umbilical cord mesenchymal stem cells could be induced to form the lumen-like structure with different diameter sizes, and umbilical cord mesenchymal stem cells could be induced to differentiate into endothelial-like cells, and have a angiogenic capacity. Figures and Tables | References | Related Articles | Metrics

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Differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells induced by liver homogenate supernatants Ma Xin, Xue Gai, Liu Jian-fang, Li Jian-li, Hou Yan-ning 2013, 17 (45):  7877-7884.  doi: 10.3969/j.issn.2095-4344.2013.45.009 Abstract ( 118 )   PDF (1227KB) ( 464 )   Save BACKGROUND: Preliminary experiments have demonstrated that rat liver homogenate supernatants can induce the morphological changes of human umbilical cord mesenchymal stem cells. However, little is known about whether the induced cells have some phenotypic and functional features of hepatocytes. OBJECTIVE: To investigate whether human umbilical cord mesenchymal stem cells have some phenotypic and functional characteristic of hepatocytes after being induced by liver homogenate supernatants. METHODS: Passage 3 human umbilical cord mesenchymal stem cells were used and divided into control group (cells were cultured in basic culture medium) and liver homogenate supernatant group (cells were cultured in liver homogenate supernatants for 3, 5, 7 days). Meanwhile, positive control group (QSG-7701 human liver cell lines) and negative control group (simple liver homogenate supernatants) were set up. The protein and mRNA level of hepatocyte markers, alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme, were detected at different time points. RESULTS AND CONCLUSION: After inducement, the stem cells of fusiform shape began to lose their sharp edges and progressively shrunk, and then they changed into hepatocyte-like cells with the morphous of triangle, polygon and anomalism shape. Compared with the control group, the protein and mRNA level of alpha-fetoprotein, cytokeratin 18 and tryptophan 2,3-dioxygenase enzyme significantly increased time dependently after inducement with liver homogenate supernatants (P < 0.01). This study demonstrated that human umbilical cord mesenchymal stem cells are able to differentiate into hepatocyte-like cells in vitro that possess some functions of liver cells. Figures and Tables | References | Related Articles | Metrics

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Effects of basic fibroblast growth factor gene transfection on the proliferation of umbilical cord mesenchymal stem cells Zheng Ying-juan, Tian Jian-chang 2013, 17 (45):  7885-7890.  doi: 10.3969/j.issn.2095-4344.2013.45.010 Abstract ( 87 )   PDF (781KB) ( 347 )   Save BACKGROUND: At present, exogenous basic fibroblast growth factor gene can be transfected into umbilical cord mesenchymal stem cells via a recombinant adeno-associated virus vector and exhibit sustained expression in transfected cells. This method can regulate cell proliferation and directed differentiation to obtain efficient long-lasting therapeutic effects. OBJECTIVE: To investigate the effects of basic fibroblast growth factor gene transfection via a recombinant adeno-associated virus vector on the proliferation and cell cycle of human umbilical cord mesenchymal stem cells cultured in vitro. METHODS: Human umbilical cord mesenchymal stem cells were cultured by the suspension culture in vitro, and were transfected by recombinant adeno-associated virus-mediated basic fibroblast growth factor gene. Cultured cells were divided into three groups: control group, basic fibroblast growth factor group, and recombinant adeno-associated virus group. Reverse transcription-PCR and western blot were used to assess the knockdown efficiency. Cellular proliferation was determined by cell growth curve and Cell Counting Kit-8 assay. The cell cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION: Compared with the other two groups, the expression of basic fibroblast growth factor mRNA and protein increased significantly, the cell growth speed was also significantly increased, the cell cycle of G0/G1 phase was decreased and cell number in S phase was increased in the basic fibroblast growth factor group after transfection. These findings suggest that the recombinant adeno-associated virus-mediated basic fibroblast growth factor gene can promote the proliferation of umbilical cord mesenchymal stem cells proliferation cultured in vitro, and also can optimize the cell culture. Figures and Tables | References | Related Articles | Metrics

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Intranasal administration of the conditioned medium of human umbilical cord-derived mesenchymal stem cells for treatment of cerebral ischemia-reperfusion injury Shen Li-ping, Wang Shuai-shuai, Dong Li-guo, Shen Xia, Hua Fang, Ye Xin-chun, Cui Gui-yun 2013, 17 (45):  7891-7897.  doi: 10.3969/j.issn.2095-4344.2013.45.011 Abstract ( 89 )   PDF (772KB) ( 446 )   Save BACKGROUND: Cytokines and neurotrophic factors secreted from human umbilical cord blood-derived mesenchymal stem cells secrete have neuroprotective effects on cerebral ischemia-reperfusion injury, but there are few reports about intranasal administration of human umbilical cord blood-derived mesenchymal stem cell conditioned medium in the treatment of stroke. OBJECTIVE: To investigate the protective effects of intranasal administration of human umbilical cord-derived mesenchymal stem cells-conditioned medium on neurologic function of rats with cerebral ischemia-reperfusion injury. METHODS: Adult rats were subjected to 2 hours of right middle cerebral artery occlusion and the human umbilical cord-derived mesenchymal stem cells were isolated from the postpartum human cord. We made the conditioned medium of human umbilical cord-derived mesenchymal stem cells. Ischemic rats were randomized and assigned to three groups and were treated by intranasal routine starting 24 hours after middle cerebral artery occlusion with: (1) saline for control group; (2) Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium for medium control group; (3) conditioned medium treatment group (10mL/kg) daily for 14 days. Behavioral tests (foot fault test, and modified Neurological Severity Score) were performed before and at 1, 7, 14 days after middle cerebral artery occlusion. RESULTS AND CONCLUSION: There was no difference in the behavioral tests among the three groups at postoperatively 1 day (P > 0.05). Compared to the control and medium control group rats, respectively, rats in the conditioned medium group significantly improved functional outcome after stroke in days 7 and 14 (P < 0.05). There was also no significant difference in functional tests between the control group and medium control group in days 7 and 14 (P > 0.05). These results suggest that human umbilical cord-derived mesenchymal stem cells-conditioned medium via intranasal administration can significantly improve neurologic functional outcome after cerebral ischemia-reperfusion injury. Figures and Tables | References | Related Articles | Metrics

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Biological characteristics of bone marrow mesenchymal stem cells from multiple myeloma patients before and after cryopreservation Wang Zhen-ling, Wang Jin-huan, Guo Qing, Zhao Zhi-gang 2013, 17 (45):  7898-7903.  doi: 10.3969/j.issn.2095-4344.2013.45.012 Abstract ( 127 )   PDF (809KB) ( 386 )   Save BACKGROUND: Bone marrow mesenchymal stem cells derived from multiple myeloma patients are characterized by pluripotential differentiation, immunoregulation and supporting hematopoiesis, but whether these features are affected after cryopreservation is unclear. OBJECTIVE: To study the biological characteristics of cryopreserved bone marrow mesenthymal stem cell derived from multiple myeloma patients. METHODS: Bone marrow mesenchymal stem cells derived from multiple myeloma patients were cryopreserved in -196 ℃ liquid nitrogen for 1 month (short term) and 12 months (long term) with Iscove’s modified Dulbecco’s medium containing 10% dimethyl sulfoxide and 40% fetal bovine serum as cryoprotectant. The viability and proliferation ability of thawed bone marrow mesenchymal stem cells were investigated. Hematopoiesis support of thawed bone marrow mesenchymal stem cells in vitro was detected by long-term bone marrow culture and the methylcellulose progenitor assay. The immunoregulatory ability of thawed mesenchymal stem cells was detected by mixed lymphocyte culture assay. RESULTS AND CONCLUSION: The cell viability was (92.9±7.5)% and (86.7±9.2)% for mesenchymal stem cells cryopreserved as long as 1 month or 12 months, respectively. Furthermore, thawed bone marrow mesenchymal stem cells possessed the ability of supporting colony forming and could significantly suppress proliferation of T lymphocytes. At last, there were no changes detected as compared with pre-cryopreserved mesenchymal stem cells in the abilities of proliferation, hematopoiesis support and immunoregulation. Cryopreservation can decrease the cell viability of bone marrow mesenchymal stem cells derived from the patients with multiple myeloma, but cannot affect the abilities of proliferation, hematopoiesis support and immunoregulation. Figures and Tables | References | Related Articles | Metrics

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Application of Accutase enzymes in the separation of spermatogonial stem cells Liu Shan-shan, Xu Li-ping, Zhu Wei-yun, Ma Ning-fang 2013, 17 (45):  7904-7910.  doi: 10.3969/j.issn.2095-4344.2013.45.013 Abstract ( 195 )   PDF (978KB) ( 509 )   Save BACKGROUND: It is reported that the cell surface antigen may be damaged by the trypsin enzyme in some extent and the cell activity may also be influenced, as a result of which, the subsequent separation and follow-up-test will be affected. Accutase enzymes possess the activities of protease and collagen enzyme and need no special termination or cleaning at the end of digestion. Another special function of accutase enzyme is to protect cell surface antigen and thus it has been widely applied in the stem cell digestion and culture. OBJECTIVE: To compare the digestive effect of accutase enzymes and trypsin enzyme in the separation and original culture of spermatogonial stem cells.  METHODS: The testes of 5-7 days male Kunming mice (n=45) were collected and primarily digested with collagenas. The suspension of digested tissue was divided into three parts with the same volume named accutases enzyme group, trypsin enzyme group and mixed enzyme group (trypsin enzyme and hyaluronidase). For the comparison of testis digestive status and the time required for the formation of single cells, the micrographs were taken at 1, 3 and 5 minutes respectively after enzyme digestion. The total number and the mortality of the single cells were estimated and compared. The leydig cells and sertoli cells were removed by differential adherent method and the remained spermatogenic cells were then treated with CD90.2 immune magnetic beads. The selected spermatogonial stem cells were subsequently labeled by glial cell line-derived neurotrophic factor receptor alpha-1 and sorted with flow cytometry. The number of spermatogonial stem cells positive for glial cell line-derived neurotrophic factor receptor alpha-1 in CD90.2+ and CD90.2- cells was compared within different groups. RESULTS AND CONCLUSION: The digestive capacities of different enzyme were different. Accutases enzyme obtained the single cell suspension more quickly than trypsin enzyme and mixed enzyme, and had the least cell masses and broken cell membrane than the other two groups. After the differential attached treatment, the highest total number of CD90+ spermatogonial stem cells and lowest cell mortality could be found in the accutases group, when compared with the other groups. The results of cell sorting by flow cytometry showed a higher rate (72.24%) of GFRα1+/ CD90+ cells in the accutases group than the trypsin group (51.16%) and mixed enzyme group (71.27%). The GFRα1+/CD90- number in the accutases group was lower (15.03%) than that of the trypsin group (18.8%) and mixed enzyme group (24.23%), respectively. The results indicate a better effect of accutases enzyme on the primary separation of spermatogonial stem cells than that of trypsin or mixed enzyme. Figures and Tables | References | Related Articles | Metrics

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Biological characteristics of endothelial progenitor cells from the peripheral blood versus umbilical cord blood Wu Hong-tao, Ma Yan, Bi Xiao-juan, Jiang Ming, Yang Kai, Wang Ning 2013, 17 (45):  7911-7917.  doi: 10.3969/j.issn.2095-4344.2013.45.014 Abstract ( 217 )   PDF (3548KB) ( 385 )   Save BACKGROUND: Endothelial progenitor cells from the peripheral blood and umbilical cord blood are an essential source to repair vascular endothelial cells damaged by various diseases. OBJECTIVE: To compare the biological characteristics of endothelial progenitor cells from peripheral blood and umbilical cord blood in vitro. METHODS: Mononuclear cells were isolated from the umbilical cord blood and peripheral blood with density gradient centrifugation method and 6% hydroxyethyl starch precipitation combined with density gradient centrifugation method, respectively. Mononuclear cells were counted. Then the cells were implanted on the culture plate pre-paved with rat tail collagen at the concentration of 1×106/cm2, and cultured in an endothelial cell culture medium for 7 days. RESULTS AND CONCLUSION: Isolated and cultured endothelial progenitor cells from the peripheral blood and umbilical cord blood had similar morphological characteristics in vitro. Under the optical microscope, with the increase of culturing days, most adherent cells were changed from round to spindle-shaped. Peripheral blood endothelial progenitor cells had cell colony formation, and spindle cells from the umbilical cord blood arranged typically in a line structure. After trypan blue staining and drawing of cell growth curves, the number of mononuclear cells and endothelial progenitor cells, survival rate and proliferative ability of endothelial progenitor cells from the peripheral blood were all lower than those from the umbilical cord blood (P < 0.05). At day 3 after incubation, the proliferation of endothelial progenitor cells from the peripheral blood and umbilical cord blood reached the peak, and then showed a decreased tendency. Flow cytometry and Immunofluorescence staining showed that endothelial progenitor cells from the peripheral blood and umbilical cord blood could express CD34, CD133, and vascular endothelial cell factor receptor 2. The endothelial progenitor cells from the peripheral blood and umbilical cord blood could swallow acetylated low density lipoprotein and be combined with ulex europaeus agglutinin Ⅰ. The results confirmed that the endothelial progenitor cells from the peripheral blood and cord blood have similar biologicla characteristics, but the proliferation ability of endothelial progenitor cells from the cord blood is higher. Figures and Tables | References | Related Articles | Metrics

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Keratinocyte serum-free medium promotes the growth of rat hair follicle stem cells Zhang Zhi-qiang, Wang Yu-jie, Muratrixat, Li Jia 2013, 17 (45):  7918-7923.  doi: 10.3969/j.issn.2095-4344.2013.45.015 Abstract ( 127 )   PDF (2351KB) ( 430 )   Save BACKGROUND: The hair follicle stem cells are usual cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10% fetal bovine serum. Recently, keratinocyte serum-free medium has been used to culture hair follicle stem cells. OBJECTIVE: To observe the effects of three different culture media on the proliferation and purity of hair follicle stem cells. METHODS: After sterling, the skin of the rat’s beard was cut off, and the vibrissa follicles were dissected under a stereocroscope. The follicles were digested by Dispase Ⅱ firstly and by the mixture of trypsin and EDTA secondly. Cell suspension was divided into three pieces based on the average number of cells and respectively cultured by keratinocyte serum-free medium, Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10% fetal bovine serum and keratinocyte serum-free medium containing 10% fetal bovine serum. Hair follicle stem cells were selected by rapid adherence on collagen type Ⅳ, and passaged. RESULTS AND CONCLUSION: Cultured hair follicle stem cells spread to the third generation, and trypan blue assay showed there was no difference in cell survival rate among the three groups (P > 0.05). Cell Counting Kit 8 assay showed that the cells grew slowly in the three groups in the first 2 days. At day 4, the hair follicle stem cells grew into the growth logarithmic phase, and the proliferative activity was ranked as follows: keratinocyte serum-free medium+10% fetal bovine serum > Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10% fetal bovine serum > keratinocyte serum-free medium with a significant difference between the three groups (P < 0.05). Flow cytometric examination showed the expression of CD34 in the keratinocyte serum-free medium+10% fetal bovine serum group was lower than that in the keratinocyte serum free medium group (P < 0.05). The expressions of CD34, CK15 and β1-integrin (CD29) in the Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10% fetal bovine serum were lower than those in the other two groups (P < 0.05). These findings indicate that we can obtain higher purity hair follicle stem cells in the keratinocyte serum-free medium+10% fetal bovine serum than in the Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10% fetal bovine serum. Figures and Tables | References | Related Articles | Metrics

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MicroRNA-1 and microRNA-499 directly reprogram cardiac myofibroblasts into cardiomyocyte-like cells Zhang Cheng1, Zhang Shao-zhong, Zhang Ya-zhou, Zhang Guo-heng, Wang Yun-ru, Dong Hong-yan, Zhang Zhong-ming 2013, 17 (45):  7924-7931.  doi: 10.3969/j.issn.2095-4344.2013.45.016 Abstract ( 265 )   PDF (3274KB) ( 657 )   Save BACKGROUND: MicroRNAs are a class of small non-coding RNA molecules. MicroRNA is involved in the regulation of gene expression, and plays an important role in promoting the proliferation and differentiation of precursor cells. Whether microRNAs can promote progenitor cells to differentiate into myocardial cells has been rarely reported. OBJECTIVE: To investigate the feasibility of direct reprogramming of neonatal mouse cardiac myofibroblasts to cardiomyocyte-like cells by specific microRNAs (microRNA-1 and microRNA-499). METHODS: Cardiac fibroblasts isolated from neonatal mice were cultured in vitro at 37 ℃ under normal oxygen, cells were used at the second passage for the following experiments, and then cultured at 37 ℃ in 3% O2 for 48 hours. Immuno?uorescent staining was used to detect expression of vimentin which is a specific marker for Cardiac fibroblasts. Immuno?uorescent staining and flow cytometry assay were employed to detect expression of α-smooth muscle actin which is a specific marker for cardiac myofibroblasts. There were four groups in the experiment: microRNA-1 group, microRNA-499 group, microRNA-1/microRNA-499 co-tranfection group, and blank control group. MicroRNA profile between cardiac myofibroblasts and myocardial cells was determined by microarray analysis via the miRCURYTM Array microarray kit. MicroRNA microarray results were validated by real-time quantitative PCR. RESULTS AND CONCLUSION: After overexpression of microRNA-1 and microRNA-499 in cardiac myofibroblasts, specific factors of heart development at different stages had different expressions; compared with the blank control group, the expression of GATA4, Tbx5, Mef-2c, α-MHC increased significantly in the remaining three groups, but Mesp1, Isl1 expression was not significantly changed). The results provide strong evidence for the capacity of microRNAs to induce expression of cardiomyocyte markers in cardiac myofibroblasts and demonstrate the potential of specific microRNAs that can direct reprogram cardiac myofibroblasts to cardiomyocytes in vitro. Figures and Tables | References | Related Articles | Metrics

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Proliferation and differentiation of mesenchymal stem cells modified with glial cell line-derived neurotrophic factor Huang Cheng, Yang Jian-dong, Feng Xin-min, Xu Wei, Li Yi-nan, Xiao Hai-xiang, Gu Jia-xiang 2013, 17 (45):  7932-7938.  doi: 10.3969/j.issn.2095-4344.2013.45.017 Abstract ( 97 )   PDF (3291KB) ( 448 )   Save BACKGROUND: Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cell viability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent. OBJECTIVE: To investigate the effect of glial cell line-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cell line-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cell line-derived neurotrophic factor. METHODS: The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes: bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. Cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cell-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified. RESULTS AND CONCLUSION: By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradually increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90% on day 3 after transfection. Cell viability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradually extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cell line-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cell line-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells. Figures and Tables | References | Related Articles | Metrics

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Transcranial magnetic stimulation promotes proliferation of endogenous neural stem cells of Parkinson’s disease model mice Gu Ping, Zhang Zhong-xia, Ma Qin-ying, Geng Yuan, Wang Yan-yong, Zhang Li-na 2013, 17 (45):  7939-7946.  doi: 10.3969/j.issn.2095-4344.2013.45.018 Abstract ( 117 )   PDF (3128KB) ( 463 )   Save BACKGROUND: Neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine can induce the clinical, biochemical and pathological characteristics similar to those observed in primary Parkinson’s disease. OBJECTIVE: To observe the effects of repetitive transcranial magnetic stimulation on the proliferation of endogenous neural stem cells of Parkinson’s disease model mice and the mood change. METHODS: A total of 72 male C57BL/6J mice were randomly divided into four groups: normal saline group, Parkinson’s disease model group (model group), sham-repetitive transcranial magnetic stimulation group (sham group) and repetitive transcranial magnetic stimulation group. The mice received 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine injection×4 to establish acute Parkinson’s disease models. The mice in the normal saline group were injected the same volume saline. And 24 hours after the last injection of 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine, the mice in the repetitive transcranial magnetic stimulation group received five trains of repetitive transcranial magnetic stimulation, 1 Hz for 25 seconds, at an intensity of 1 Tesla daily for 1, 3, 7 consecutive days. Sham group mice were not exposed to the magnetic field. No treatment was performed in the mice of model group. The mood change was evaluated using the elevated-plus maze testing before and after repetitive transcranial magnetic stimulation treatment. The change in expression of nestin in the subventricular zone was observed by using immunohistochemical technique. RESULTS AND CONCLUSION: (1) Elevated-plus maze testing: There was no statistical significance about percentage of opening arm time accounting for total time among groups and at different time points in each group, but after stimulation, the percentage of opening arm time accounting for total time showed a declined tendency. (2) The results of nestin immunohistochemical staining: Compared to the normal saline group, the number of nestin-positive cells of the model group was increased at days 3 and 7, and there was no statistical significance in the number of nestin-positive cells between model group and sham group; Compared to the sham group and model group, the number of nestin-positive cells of repetitive transcranial magnetic stimulation group were evidently increased; The proliferation of endogenous neural stem cells was time-dependent, endogenous neural stem cells exhibited outward migration gradually along the certain way, and some cells were able to migrate to the corpus callosum at day 3, and even to the cerebral cortex at day 7. Repetitive transcranial magnetic stimulation can promote the endogenous neural stem cells in a time-depended manner. Figures and Tables | References | Related Articles | Metrics

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In vitro cultivation, identification and osteoinduction of adult bone marrow mesenchymal stem cells Zhang Ming-jie, Zhang Qing-wen, He Wei, Chen Zhen-qiu, Ou Zhi-xue, Jia Xiao-jun 2013, 17 (45):  7947-7953.  doi: 10.3969/j.issn.2095-4344.2013.45.019 Abstract ( 85 )   PDF (4387KB) ( 373 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have the potential of self-proliferation and multi-directional differentiation, while mesenchymal stem cells are few in adult bone marrow. In vitro purification, amplification and osteoinduction are very important for the research of bone tissue engineering. OBJECTIVE: To establish a simple and reliable in vitro cultivation and identification system of adult bone marrow mesenchymal stem cells, and to induce the mesenchymal stem cells to differentiate into osteoblasts. METHODS: Bone marrow were extracted from adult anterior superior iliac, the density gradient centrifugation and adhesion method were used to isolate, purify, culture and amplify the bone marrow mesenchymal stem cells. Osteogenic medium was prepared by mixing appropriate amount of dexamethasone, β-glycerophosphate and ascorbic acid C. The cells were divided into osteoinduction group and blank control group for observation. RESULTS AND CONCLUSION: Adult bone marrow mesenchymal stem cells were in typical long spindle-shape. The cells grew into rapid proliferation phase at 8-11 days and the growth curve was S-shape. CD44 and CD90 were in positive expression, while CD34 and CD45 were negative. The alkaline phosphatase activity was increased with culturing time prolonging, and reached the summit at the 12th day. The alkaline phosphatase activities of osteoinduction group were higher than those in the blank control group at different time points. These results suggested that in vitro cultivation, identification and osteoinduction system could obtain mesenchymal stem cells with high purity and good osteogenic differentiation capacity. Figures and Tables | References | Related Articles | Metrics

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Stem cell therapy for ischemic stroke Yuan Zhi-jun, Li Xiao-gang 2013, 17 (45):  7954-7960.  doi: 10.3969/j.issn.2095-4344.2013.45.020 Abstract ( 104 )   PDF (605KB) ( 458 )   Save BACKGROUND: Stem cells are a kind of cells characterized as species diversity, self-replication and renewal ability, multiple differentiation potential and high proliferation potential. Then, the stem cell treatment for ischemic brain injury would be of great benefit. Stem cell therapy provides a new way for the treatment of ischemic stroke, but the mechanism is still unclear. OBJECTIVE: To describe the types of stem cells and review the mechanism underlying stem cell treatment for ischemic stroke. METHODS: The first author retrieved PubMed database, Chinese Journal Full-text Database for articles related to stem cell classification and effectiveness, safety and mechanism of stem cell therapy for ischemic stroke published from January 1992 to September 2012. The key words were “stem cells, brain ischemic stroke, transplantation, treatment” in English and Chinese, respectively. A total of 168 literatures were retrieved, and 61 articles met the inclusion criteria. RESULTS AND CONCLUSION: Stem cell therapy for ischemic stroke has shown a promising prospect though it is staged in the period of animal models. Stem cell transplantation for promoting functional recovery in the treatment of stroke has been completed in the clinical phase I or phase II trials. Stem cell transplantation for ischemic stroke appears to have no adverse reactions and to promote functional recovery. Main difficulties in stem cell transplantation for treatment of ischemic stroke include sources of stem cells, transplantation approach, stem cell survival in the host body, stem cell integration with the host brain, therapeutic effectiveness and security. Based on the acquired results from the mechanism research and clinical trials, how to safely and quickly apply the stem cell therapy from the experiments to the clinic still needs to work. Figures and Tables | References | Related Articles | Metrics

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Deer antler: A stem cell-based organ regeneration research model Chu Wen-hui, Wang Da-tao, Lu Xiao-ping, Li Chun-yi 2013, 17 (45):  7961-7967.  doi: 10.3969/j.issn.2095-4344.2013.45.021 Abstract ( 86 )   PDF (708KB) ( 672 )   Save BACKGROUND: Deer antlers are the unique mammalian organs which can periodically regenerate, and the process is known as a stem cell-based event. Exploring the underlying mechanism of deer antler regeneration and indentifying the functional role of stem cell in mammalian organ regeneration are of great importance to regenerative biology and regenerative medicine. OBJECTIVE: To review the relevant literatures of the research progress in antler regeneration, as well as effects of stem cells and cytokines on antler regeneration. METHODS: A computer-based online search of PubMed (1994-01/2012-10) was performed for acquiring the articles in English by using the key words of “deer antler; antler regeneration; stem cell. In addition, manual search was also performed for those literatures that cannot be readily obtained from internet search. Articles concerning antler regeneration histology, morphology, antler stem cells and micro-environmental studies, and related cytokines. Repetitive studies or articles that are unrelated to the criteria set for the article were excluded. RESULTS AND CONCLUSION: A total of 87 articles were obtained and finally 31 articles were selected. The key tissue types for antler regeneration are antlerogenic periosteum and pedicle periosteum, the cells within which are known as antler stem cells. The covering skin of antlerogenic periosteum and pedicle periosteum constitutes the functional niche for antler stem cells. Numerous cytokines are involved in the process of antler fast growing and full regeneration, including insulin-like growth factor, sex hormones, human epidermal growth factor, and vascular endothelial growth factor. It is vitally important to identify the interacting molecules between the antler stem cells and their niche cell types, and to define the role of each molecule that plays in antler regeneration, which will greatly advance our knowledge of the stem cell-based mammalian organ regeneration. Figures and Tables | References | Related Articles | Metrics

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Mechanism underlying mesenchymal stem cells to regulate autoimmune thrombosis of systemic lupus erythematosus Ren Min-min, Gu Jian 2013, 17 (45):  7968-7974.  doi: 10.3969/j.issn.2095-4344.2013.45.022 Abstract ( 79 )   PDF (886KB) ( 426 )   Save BACKGROUND: Systemic lupus erythematosus is a complex autoimmune disease of unknown origin affecting virtually every organ in the human body. It is characterized by abnormal activation of T, B lymphocytes. While aberrant T cells provide help to autoreactive B cells, the autoreactive B cells produce a variety of pathological autoantibodies and immune complex deposition, which could infiltrate the small blood vessels directly or deposit in the vessel wall, thereby causing inflammatory necrosis of the vessel walls. These changes will narrow the vascular lumen, and promote thrombosis, leading to local tissue ischemia and dysfunction. Mesenchymal stem cells can regulate the immune disorders, play an anti-inflammatory role, and repair the immune thrombosis. OBJECTIVE: To summarize the pathogenesis of immune thrombosis in systemic lupus erythematosus, as well as to introduce the mechanism of mesenchymal stem cell therapy for immunity thrombosis. METHODS: We searched the PubMed database, Springlink database, ScienceDirect database and HighWire database from January 1990 to June 2013 with the key words of “systemic lupus erythematosus, mesenchymal stem cell, thrombosis, T cells, B cells” in English. An online search of CNKI database, Wanfang database, VIP database from January 1990 to June 2013 was also conducted with the key words of “systemic lupus erythematosus, mesenchymal stem cell, thrombosis, T cells, B cells” in Chinese. A total of 267 literatures were screened out, and 48 documents were included in the review based on the inclusion and exclusion criteria. RESULTS AND CONCLUSION: Mesenchymal stem cells are multipotential nonhematopoietic progenitor cells capable of multi-directional differentiating into various cell types. Mesenchymal stem cells can suppress T-lymphocyte activation and proliferation, and inhibit secretion of interferon-γ, interleukin-4. Mesenchymal stem cells can also inhibit the proliferation of B cells and the secretion of pathogenic immunoglobulin IgM, IgG, IgA. Thus, mesenchymal stem cells can regulate immune homeostasis and reduce the deposition of autoantibodies and immune complexes, thereby protecting the blood vessels from injury, reducing the secretion of inflammatory cytokines, balancing the coagulation-anticoagulation, and decreasing thrombosis in systemic lupus erythematosus. Figures and Tables | References | Related Articles | Metrics

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Umbilical cord blood mononuclear cell transplantation for treatment of decompensated cirrhosis Xu Yu-qin, Shi Wen-ming, Li Jin-shun, Xu Jian-guo, Li Hong-wen, Hu Xiang 2013, 17 (45):  7975-7980.  doi: 10.3969/j.issn.2095-4344.2013.45.023 Abstract ( 104 )   PDF (945KB) ( 410 )   Save BACKGROUND: Orthotopic liver transplantation is the most effective therapy for the treatment of end-stage liver diseases, but the lack of donor source, immune rejection, and repeated infections limit its application. Stem cell transplantation technology provides a new idea for the treatment of end-stage liver diseases. A variety of methods have been confirmed to successfully induce umbilical cord blood mesenchymal stem cells converted into liver cells in vitro. OBJECTIVE: To explore the clinical efficacy and feasibility of human umbilical cord blood mononuclear cells transplantation in the treatment of decompensated cirrhosis. METHODS: Twenty-three patients with decompensated cirrhosis received allogeneic human umbilical cord blood mononuclear cell transplantation. Serum alanine aminotransferase, albumin, cholinesterase, total bilirubin and prothrombin time were detected at post-transplantation weeks 2, 4, 8 and 24. Improvement in clinical signs and symptoms as well as adverse reactions was observed. RESULTS AND CONCLUSION: Liver function had no changes at 2 weeks after human umbilical cord blood mononuclear cell transplantation (P > 0.05). At 4 weeks after cell transplantation, serum alanine aminotransferase was improved significantly (P < 0.05), but the other indexes still had no changes. Until 12 weeks after cell transplantation, there were significant improvements in all the liver function indicators (P < 0.05) and the liver stiffness (P < 0.05). By the end of 24 weeks, all the test results were improved significantly (P < 0.01). Clinical symptoms were alleviated, including fatigue improvement in 20 cases (87%), improved appetite in 21 cases (91%), and relieved ascites in 19 cases (83%). No severe adverse reactions were found during the transplantation and 24-week follow-up. These findings suggest that human umbilical cord blood mononuclear cells transplantation is effective and safe for the treatment of decompensated cirrhosis, which can be considered as a clinical therapy for patients with advanced cirrhosis. Figures and Tables | References | Related Articles | Metrics