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03 December 2010, Volume 14 Issue 49 Previous Issue    Next Issue

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Effects of neotype bone morphogenetic protein-7 peptide on biological behavior of bone marrow stromal stem cells Feng Bo, Hu Dong-xu, Zhang Yang-de, Wu Yu-chi 2010, 14 (49):  9121.  doi: 10.3969/j.issn.1673-8225.2010.49.001 Abstract ( 328 )   PDF (430KB) ( 402 )   Save BACKGROUND: The study of bone morphogenetic protein-7 (BMP-7) mainly focused on its application to tissue engineering. However, BMP-7 has strong bone induction activity and affects growth, development and physiological function of animals’ reproductive system. How to fully exert its bone induction activity and to avoid other biological effects is a difficulty in bone tissue engineering field. OBJECTIVE: To explore the effect of BMP-7 peptide on proliferation, osteogenic differentiation and mineralization of bone marrow stromal stem cells (BMSCs). METHODS: BMSCs were harvested from rats and identified with flow cytometry. The 3rd generation of BMSCs was divided into experimental and control groups. The medium of experimental group was supplemented with 100 mg/L BMP-7 peptide. The medium of control group was not supplemented with BMP-7 peptide. After 2, 4, 6, 8, and 10 days, the proliferation of BMSCs was analyzed by MTT assay. After 7 and 14 days, the calcium contents and alkaline phosphatase activities were measured to assess the differentiation of BMSCs towards osteoblasts. The mineralization of BMSCs was determined by calcein staining. RESULTS AND CONCLUSION: BMSCs grew well in primary culture and the purity of BMSCs was 91.5%. There was no significant difference in proliferation between experimental and control groups (P > 0.05). On days 7 and 14, calcium contents and alkaline phosphatase activities in the experimental group were significantly higher than the control group (P < 0.05). The mineralization level of BMSCs in the experimental group was significant higher than in the control group. Results suggest that BMP-7 peptide has no influence on the proliferation of BMSCs, but it can promote osteogenic differentiation and mineralization of BMSCs significantly. Related Articles | Metrics

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Different incubation densities of bone marrow mesenchymal stem cells and bone matrix gelatin complex in repair of radius defects in rats Tian Zhi-feng, Qin Shu-jian, Zhang Xiao-ling, Li Zhan-sheng 2010, 14 (49):  9126.  doi: 10.3969/j.issn.1673-8225.2010.49.002 Abstract ( 368 )   PDF (602KB) ( 399 )   Save BACKGROUND: When the complex was made, the seed cell’s vaccination density will affect the ability of the complex, but the appropriate incubation density was not still unified. OBJECTIVE: To construct bone marrow mesenchymal stem cells (BMSCs)/bone matrix gelatin (BMG) complex and to observe the optimal cell density of BMSCs/BMG complex. METHODS: BMSCs from Sprague Dawley rats were cultured in vitro. BMSCs at different cell densities were incubated on the heterogenic BMG. BMSCs/BMG complex was constructed, and the adherent growth of cells on the BMG was observed using scanning electron microscope. A bone defect model (5 mm in length) was created at bilateral radius in each Sprague Dawley rat. This study was divided into four groups. BMSCs/BMG complex was used to repair bone defect. The seeding density was respectively 1×108/L, 5×108/L, 1×109/L and 5×109/L. At 2, 4, 8 and 12 weeks, the samples were obtained in each group. Gross observation on the activity of operated limb, radiographic and histological examination and immunohistochemical analysis were performed to observe the repair condition of radius defect after surgery.  RESULTS AND CONCLUSION: After 24 hours of BMSCs/BMG complex culture, the adherent rate of cells on the BMG increased with increased cell incubation density. When the densities surpassed 1×109/L, and the maximum adhesive rate was (76.00±2.94)%. Increased cell densities showed decreased adhesive rate. At day 7, results of scanning electron microscope exhibited cells adhered on the material. X-ray radiology score showed the score was greater at 8 and 12 weeks compared with that at 2 weeks in the same group (P < 0.01). No significant difference was determined at 4, 8 and 12 weeks in the 1×109/L and 5×109/L groups (P > 0.05), and the score was greater than other two groups (P < 0.01). Histological score results have demonstrated that the score was higher at 8 and 12 weeks than that at 2 weeks in the same group (P < 0.01). Results suggest that when constructing the BMSCs/BMG complex in vitro, the optimal BMSCs seeding density was 1×109/L. Related Articles | Metrics

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Gene expression during induced differentiation of bone marrow mesenchymal stem cells into osteoblasts Hu Yang, Ma Ying, He Hui-yu, Sheng Lei, Abulizi•Abudula, Aerziguli 2010, 14 (49):  9131.  doi: 10.3969/j.issn.1673-8225.2010.49.003 Abstract ( 578 )   PDF (540KB) ( 533 )   Save BACKGROUND: At present, the gene expression mode of bone marrow mesenchymal stem cells (BMSCs) differentiation into osteoblasts remains unclear. OBJECTIVE: To observe gene expression of alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin during BMSCs differentiation into osteoblasts, and to verify whether differentiation of BMSCs into osteoblasts is mature. METHODS: The BMSCs were aspirated from the femoral bone of 2-month old New Zealand rabbits and cultured by all bone marrow adherence method, and induced to differentiate into osteoblasts in the medium supplemented with DMEM/F12, dexamethason 1×10-8 mmol/L, β-glycerophosphate sodium 0.01 mol/L and vitamin C 0.05 g/L. The gene expression of alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin in the first and second passages of BMSCs were examined by reverse transcription-polymerase chain reaction. BMSCs surface antigen CD44 was identified. RESULTS AND CONCLUSION: Following induced culture in mineralized media, the first and second passages of BMSCs expressed alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin gene in some phases. Positive rate of CD44 in the first passage of BMSCs was 44.4%. These indicate that rabbit BMSCs gradually differentiated into osteoblasts following in vitro mineralization. The first and second passages of BMSCs expressed alkaline phosphatase, osteopontin, type Ⅰcollagen, basic fibroblast growth factor and osteocalcin in some phases. These cells have had the characteristics of osteoblasts and provided experimental evidences for revealing the mechanisms of gene expression of BMSCs differentiation into osteoblasts. Related Articles | Metrics

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Directional differentiation of human bone marrow mesenchymal stem cells into neuron-like cells in vitro Wang Ying, Zhao Hong-chang, Zhao Wen-jing, Ye Dong-xia, Li Jing, Luo Su 2010, 14 (49):  9138.  doi: 10.3969/j.issn.1673-8225.2010.49.004 Abstract ( 365 )   PDF (351KB) ( 384 )   Save BACKGROUND: In vitro induced bone marrow mesenchymal stem cells (BMSCs) differentiated into neuron-like cells mainly by polypeptide preparations such as nerve growth factor. Pure chemical inducer is not commonly found. OBJECTIVE: To establish a system for isolation and culture of BMSCs and induce it directional differentiation into neuron-like cells in vitro. METHODS: BMSCs were isolated, purified and identified by density gradient centrifugation, adherent culture and digestion time control. BMSCs were induced into neuron-like cells by β-mercaptoethanol and dimethyl sulfoxide. The morphology change was observed. The differentiation ratio analysis of differentiated neuron-like cells was detected by Nissl staining and immunochemical of neuron specific enolase and neurofilament-200. RESULTS AND CONCLUSION: Separated BMSCs were fibroblast-like cells and they had several nucleoluses. BMSCs could differentiate into neuron-like cells, with the presence of long axons and dendrite-like processes after being induced by β-mercaptoethanol and dimethyl sulfoxide. Both Nissl’s body, neurofilament -200 and neuron specific enolase of neurons after induction were positive and the positive rates of neuron specific enolase and neurofilament-200 were (85.6 ±6.7)% and (73.2 ± 5.6)%, respectively. Results demonstrated that density gradient centrifugation, adherent culture and digestion time control can successfully isolate and culture human BMSCs. Human BMSCs can differentiate into neuron-like cells induced by β-mercaptoethanol and dimethyl sulfoxide in vitro. Related Articles | Metrics

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Cultivation optimization and multi-directional differentiation potential of adult bone marrow mesenchymal stem cells Wang Qing-de, Lian Hong-kai, Wang Chun-ping, Wang Chun-li, Bai Yu 2010, 14 (49):  9142.  doi: 10.3969/j.issn.1673-8225.2010.49.005 Abstract ( 233 )   PDF (358KB) ( 323 )   Save BACKGROUND: The number of adult bone marrow mesenchymal stem cells (BMSCs) is few. To apply BMSCs to tissue engineering and cell therapy, we should establish mature, simple and effective isolating culture amplification system. OBJECTIVE: To establish a method of isolation and cultivation in vitro of adult BMSCs, and to observe the results of BMSCs differentiating into the adipocytes, osteoblasts and chondrocytes. METHODS: Adult BMSCs were respectively isolated and cultivated by density gradient centrifugation and whole bone marrow culture. Cell morphology was observed under an optical microscope to draw growth curves. The effects of different culture methods on BMSCs were compared. CD34, CD44 and CD105 expressions were detected by immunofluorescence. Adult BMSCs were induced with different media. Differentiations of BMSCs into adipocytes, osteoblasts and chondrocytes were measured using Oil Red O staining, alizarin red staining and alcian blue staining. RESULTS AND CONCLUSION: By the method of whole bone marrow culture, the characteristics of proliferation and growth of BMSCs were superior to that by the method of density gradient centrifugation. Immunophenotypically, this cell population was found to be highly positive for CD44 and CD105, while negative for CD34. Following induced culture, Oil Red O staining, alizarin red staining and alcian blue staining were positive, which verified that BMSCs harvested by the method of whole bone marrow culture gained more capacity of growth and proliferation, and BMSCs could be induced to differentiate into osteoblasts, adipocytes and chondrocytes. Related Articles | Metrics

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Comparison of biological characteristics of rat bone marrow stromal stem cells at different ages 2010, 14 (49):  9147.  doi: 10.3969/j.issn.1673-8225.2010.49.006 Abstract ( 285 )   PDF (311KB) ( 373 )   Save BACKGROUND: Bone marrow stromal stem cells (BMSCs) have multiple differentiation abilities. BMSCs are easily obtaining, have strong proliferative ability and can do autologous transplantation. Therefore, it is an ideal seed cells in tissue engineering. Few studies have addressed effects of age on biological characteristics of BMSCs. OBJECTIVE: To compare differences in biological characteristics of BMSCs from rats at different ages, and to provide experimental evidence for clinical cell transplantation. METHODS: BMSCs were obtained from Sprague-Dawley rats. SD rats were divided into three groups according to the source of cells, namely childhood group, adult group, gerontism group. The femur at both ends was cut under sterile conditions. Bone marrow cavity was washed using DMEM medium containing 10% fetal bovine serum. Bone marrow fluid was collected. Single cell suspension was made, and placed in culture flask after filtration through 200 meshes. The second passage of cells was obtained. CD44, CD29, CD90 protein expression was determined using immunofluorescence staining. Morphology and adhesion rate of BMSCs were observed. The cell growth curve was drawn by cells counting. Activity of BMSCs was detected using MTT assay. RESULTS AND CONCLUSION: The cells were positive for the CD44, CD29 and CD90 immunostaining. The adhesion rate of childhood group was faster than adult and gerontism groups as well as the proliferation rate (P < 0.05). MTT assay showed that the viability of childhood group was significantly stronger than the other two groups (P < 0.05). Results indicated that BMSCs from the young rats are the ideal tissue engineering cells, for the characters of dynamic and fast amplification in a short time. Related Articles | Metrics

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Proliferation of bone marrow mesenchymal stem cells under different serum culture conditions Wu Gang, He Hui, Cao Yue-cheng, Chen Xu-chun, Yang Lei, Fu Da-zhi 2010, 14 (49):  9151.  doi: 10.3969/j.issn.1673-8225.2010.49.007 Abstract ( 409 )   PDF (338KB) ( 666 )   Save BACKGROUND: There are many studies concerning bone marrow mesenchymal stem cells for culturing methods, and fetal bovine serum (FBS)/ fetal calf serum (FCS) are used widely. However, there is few study about cord blood serum (CBS) culture method on bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To explore the effects of different serum culture method on the proliferation of BMSCs. METHODS: Ficoll density gradient centrifugation method was used to isolate and purify BMSCs that were incubated in L-DMEM containing CBS and FBS and serum-free media. Morphology and proliferation of BMSCs were observed. CD34, CD44, CD45 and CD105 were identified using flow cytometry. RESULTS AND CONCLUSION: CBS group cells appeared smaller, more spindle-shaped with fewer cytoplasmic processes than the cells cultured with FBS. CBS and FBS could promote cell growth and proliferation, but BMSCs in CBS were active, but no proliferation was determined in serum-free media. Flow cytometry showed that BMSCs were negative for CD34, CD45, but positive for CD105 and CD44. Surface marker on BMSCs did not significantly change following CBS amplification. Related Articles | Metrics

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Pravastatin effects on steroid-induced differentiation of bone marrow stromal cells Wen Guo-hong, Xiao Zeng-ming 2010, 14 (49):  9155.  doi: 10.3969/j.issn.1673-8225.2010.49.008 Abstract ( 261 )   PDF (259KB) ( 284 )   Save BACKGROUND: Pravastatin as a hypolipidemic drug is widely used in clinic. With the deep research of this kind of drugs, it has effects on bone formation. Whether pravastatin can inhibit the differentiation of hormone-induced bone marrow stromal cells into adipocytes remains poorly understood. OBJECTIVE: On the base of steroid-induced adipogenesis in bone marrow stromal cells, to observe the influence of pravastatin and comment on the mechanism of prevention and treatment of steroid-induced necrosis of femoral head. METHODS: Bone marrow of bilateral femur of young rabbits was obtained, and bone marrow stromal cells were isolated by cell adherence. The osteoprogenitor cells were treated with dexamethasone which acted as adipocyte differentiation inducer and various mass concentrations of pravastatin. The cells in culture were stained on day 21 following culture. The number of adipocytes was observed under a light microscope, and counted on a pathological image analyser. Cultured cells received immunohistochemical staining on day 12. Activities of alkaline phosphatase of the cells were measured. RESULTS AND CONCLUSION: On day 21 following culture, the number of adipocytes in each group was reduced with the increased dose of pravastatin, in a dose dependent fashion. In addition, on day 21 following treatment of dexamethasone and different mass concentrations of pravastatin, immunohistochemical method confirmed that the activities of alkaline phosphatase were significantly increased. Results suggest that dexamethasone can induce the differentiation of bone marrow stromal cells into adipocytes, and reduce osteoblast differentiation. Pravastatin can suppress the differentiation of bone marrow stromal cells into adipocytes, and contribute to osteoblast differentiation. Related Articles | Metrics

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Effect of bone marrow mesenchymal stem cell transplantation on alveolar fluid clearance in acute lung injury rats Zhang Jun-zhi, Wang Ping, Yao Shang-long, Liu Tang-hua, Gui Ping 2010, 14 (49):  9159.  doi: 10.3969/j.issn.1673-8225.2010.49.009 Abstract ( 293 )   PDF (359KB) ( 324 )   Save BACKGROUND: The increase of alveolar fluid is an important pathophysiological change of acute lung injury. Repairing the damaged alveolar-capillary membrane barrier is an effective way of alleviating pulmonary edema. OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) transplantation on alveolar fluid clearance in endotoxin-induced acute lung injury rats. METHODS: Rat BMSCs were isolated and cultured using the modified Peister method. At the third passage, the BMSCs were made into single cell suspension. The rats were randomly divided into three groups. In the acute lung injury and cell transplantation groups, endotoxin was injected into the abdomen to establish models of acute lung injuny. One hour following model establishment, the intervention group, by caudal vein injection, was injected with BMSC suspension of 0.5 mL (cell count 1 × 106), and the control group and damage group were given the equal volume of normal saline. For each group, the rate of alveolar fluid clearance was measured, the lung water content level was assayed, and the pathological changes of lung tissue were observed after tail vein injection at 6, 24 and 48 hours respectively. RESULTS AND CONCLUSION: The lung water content level assay showed that the wet to dry ratio for lung injury group was significantly higher (P < 0.05) at 24 hours, compared with the normal control group and cell transplantation group. Compared with the lung injury group, the wet to dry ratio of the cell transplantation group was significantly lower (P < 0.05) at various time points. For the cell transplantation group, the rate of alveolar fluid clearance was significantly higher at various time points, compared with the lung injury group (P < 0.05). By hematoxylin-eosin staining microscopy, the lung injury group showed capillary congestion and alveolar infiltration of inflammatory cells at 6 hours, which aggravated at 24 hours. The cell transplantation group showed that at 6 hours the alveolar space infiltration of inflammatory cells was better, compared with the lung injury group, which continued to get better at 24 hours and become basically close to the normal control group at 48 hours. This indicated that BMSC transplantation may be able to promote the alveolar fluid clearance in acute lung injury by repairing damaged alveolar-capillary membrane barrier. Related Articles | Metrics

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Intraarticular injection of bone marrow mesenchymal stem cells for treatment of early osteoarthritis detected by magnetic resonance Liu Zhao, Cui Yan-an, Huang Hai-qing, Zhou Zhong-heng, Zhuang Chao 2010, 14 (49):  9163.  doi: 10.3969/j.issn.1673-8225.2010.49.010 Abstract ( 301 )   PDF (315KB) ( 403 )   Save BACKGROUND: Authors have assessed the FS-3D-FISP sequence for diagnosis of osteoarthritis. This sequence has a good consistency of the grade and pathology of early osteoarthritis. OBJECTIVE: To observe the thickness and signal changes by magnetic resonance (MR) in the treatment of on early osteoarthritis in rabbits using bone marrow mesenchymal stem cells (BMSCs) transplantation. METHODS: New Zealand white rabbits were randomly and equally divided into a control group, osteoarthritis group and BMSCs group. Injection of BMSCs was given to the left knee joint of rabbits in the BMSCs group on week 4 after arthritis induction. MR of knee joints was done at 2 weeks, 1, 2 and 3 months after the injection; three rabbits were randomly selected every time from each group. Using the sequence of FS-3D-FISP, the thickness of articular cartilage was measured and the T2-FSE double-eho sequences were utilized to measure the changes of T2 value after treatment. Pathohistological changes were observed as well as Mankin scores. The effect of the osteoarthritis in rabbit was evaluated. RESULTS AND CONCLUSION: Compared with osteoarthritis group, the thickness of articular cartilage got close to normal; T2 value reduced, and without arthroedema shown in magnetic resonance imaging. Results indicated that the thickness and signal changes of the articular cartilage on MR can be used to evaluate the changes in osteoarthritis pre- and post-treatment. Related Articles | Metrics

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Ultrastructure of injured spinal cord following olfactory ensheathing cells transplantation Liu Xiao-yang, Sun Jian-min, Cui Xin-gang, Jiang Zhen-song 2010, 14 (49):  9167.  doi: 10.3969/j.issn.1673-8225.2010.49.011 Abstract ( 235 )   PDF (437KB) ( 358 )   Save BACKGROUND: Neural function automatically recovers a little after spinal cord injury. Because of the capacity of myelination belonging to both peripheral and central glial cells, olfactory ensheathing cells (OECs) become the most promising cells for repairing damaged nerves. The histological and ultrastructural changes of spinal cord after OECs grafting may help to explain the mechanism of neural restoration. OBJECTIVE: To assess the effect of transplantation of OECs originating from the olfactory bulbs to repair spinal cord injury, and explore the influence of grafted OECs on histology and ultrastructure of neurons and axons. METHODS: Wistar rat models were randomly divided into three groups: the control group (group A, no treatment), DMEM/F12 group (group B, administrated with DMEM/F12 medium) and the OECs group (group C, treated with OEC suspension). The Basso, Beattie and Bresnahan locomotor rating scale (BBB) was observed each week. Histological changes and immunohistochemical staining were observed 8 weeks later. Reparation of spinal cord injury was assessed. Effects of OEC transplantation on spinal cord tissue and ultrastructure were observed. RESULTS AND CONCLUSION: The posterior limb motor function was recovered in three groups. The BBB grade of groups    C was higher than that of group A and group B, especially in the last 4 weeks post operation. Histology revealed that both stumps were connected by nerve fibers through the injury site in group C. Less damage of nerve fibers and neurons at the vicinity of damage site was observed in group C than that in group A and group B. The percentage of caspsase-3 positive cells in group   C was smaller than that in group A and group B. The ultrastructure showed that morphology of fiber and neuron in group C was better than that in groups A and B. Results suggest that transplantation of OECs has significant promotory effects on restoration of spinal cord injury in the rat, and can improve part function of nerve fibers and shows neuroprotection on neurons and nerve fibers. Related Articles | Metrics

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Autologous bone marrow mesenchymal stem cell transplantation via lumbar puncture in the treatment of cerebral infarction sequelae in 6 cases Jiao Hong-liang, Guan Fang-xia, Yang Bo, Li Jian-bin, Zhan San-hua, Du Ying, Hu Xiang, Zhao Lin-na, Han Xiao-gai 2010, 14 (49):  9172.  doi: 10.3969/j.issn.1673-8225.2010.49.012 Abstract ( 398 )   PDF (344KB) ( 374 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have multi-directional differentiation potential, and are easily, rapidly expanded, low immunogenicity, avoid immunological rejection in the transplant process, are ideal seed cells in the treatment of neurological diseases such as cerebral infarction. OBJECTIVE: To explore the safety, feasibility and efficacy of autologous bone marrow mesenchymal stem cell after culture transplantation via lumbar puncture in treatment of cerebral infarction. METHODS: A total of 6 cases of sequelae of cerebral infarction received autologous bone marrow mesenchymal stem cell transplantation. Autologous bone marrow mononuclear cells were collected by bone puncture. Bone marrow mesenchymal stem cells in vitro cultured 4 days after transplantation by lumbar puncture in the subarachnoid space. The efficacy and complications were observed after transplantation. RESULTS AND CONCLUSION: Numbers of BMSCs were significantly increased after culture. There were significant differences in NIHSS scores and muscle strength of patients before and after operation (P < 0.05) and the overall improvement of symptoms were improved significantly. After transplantation, there were no serious adverse events. The results suggest that autologous bone marrow mesenchymal stem cell transplantation can significantly improve symptoms in patients with cerebral infarction sequelae; treatment is safe, effective and low side effects. Related Articles | Metrics

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Migration and differentiation of neural stem cells implanted into the ventricle in neonatal rats with periventricular leukomalacia He Yue-qiu, Chen Hui-jin, Qian Long-hua, Chen Guan-yi 2010, 14 (49):  9176.  doi: 10.3969/j.issn.1673-8225.2010.49.013 Abstract ( 286 )   PDF (339KB) ( 352 )   Save BACKGROUND: The abilities of proliferation, migration, differentiation, axon formation and myelinization of transplanted neural stem cells (NSCs) were greater in the brain of new-born animals compared with adult animals. OBJECTIVE: To observe the migration and differentiation of exogenous NSCs transplanted into ventricle in neonatal rats with periventricular leukomalacia (PVL), and to explore the feasibility of NSCs transplantation for the treatment of PVL. METHODS: The intraventricular transplantation of exdogenous NSCs labeled with PKH26 fluorescein was undertaken in two-day-old neonatal rats with PVL at 72 hours after operation. The ventricular puncture point under stereotaxis instrument was: AP=1.5 mm, ML=-2 mm, DV=1.5 mm. The cell density implanted was 5×1010/L; the volume dose was 2 μL and the velocity was 0.5 μL/min. NSCs implanted was dynamically observed under laser scanning confocal microscope and immunefluorescence analysis was undertaken for the confirmation of differential cells originated from NSCs at 1, 2, 3, 7, 14 and 21 days after transplantation, respectively. RESULTS AND CONCLUSION: Laser confocal microscopy was used to observe transplanted NSCs, and confirmed that exogenous NSCs implanted through ventricle hold a favorable ability of migration. The most exogenous NSCs migrated into periventricular area and distributed in the serious injured region of white matter within three days after transplantation. Since two weeks after transplantation, NSCs implanted had differentiated mainly into OLs precursors, and partly into neurons or astrocytes in the periventricular area. These suggested that intraventricular transplantation of NSCs is with a great potential feasibility for the treatment of PVL. Related Articles | Metrics

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Changes in motor function and evoked potentials of rats with acute spinal cord injury following olfactory ensheathing cells transplantation combined with Methyprednisolone Wang Bin, He Xi-jing, Li Qiang, Wang Wei-xiong, Shen Bo 2010, 14 (49):  9181.  doi: 10.3969/j.issn.1673-8225.2010.49.014 Abstract ( 291 )   PDF (279KB) ( 353 )   Save BACKGROUND: Olfactory ensheathing cells (OECs) transplantation and Methyprednisolone (MP) are identified as two very promising methods in treating spinal cord injury (SCI). However, there are few reports about OECs combined with MP in treating SCI, and the treatment results are different. OBJECTIVE: To investigate the behavioral score and electrophysiological effects of OECs transplantation and MP on the repair of acute spinal cord injury and whether there is a synergistic effect between OECs transplantation and MP. METHODS: Acute spinal cord injury model (T10) was established by NYU impactor. All rats were randomly divided into 6 groups: control group, SCI group, DF12 group, OECs transplantation group, MP group, and OECs + MP group. Rats were injected with OECs, MP, OECs + MP, DF12 serum-free medium and saline, respectively after operation. At 8 weeks following operation, motion evoked potential (MEP) and somatosensory evoked potential (SEP) tests were performed. Changes in motor function before and after operation were estimated by Basso-Beattie-Bresnahan (BBB) score. RESULTS AND CONCLUSION: Compared with the SCI and DF12 groups, BBB score was increased, the latency period in SEP and MEP was decreased and wave amplitude was increased in the OECs, MP, and OECs + MP groups at 8 weeks following operation. Significant difference was detected (P < 0.05). The latency period in SEP and MEP was decreased and wave amplitude was increased in the OECs + MP group compared with the OECs group and MP group, and significant difference was detected (P < 0.05). OECs transplantation and MP alone can significantly promote recovery of motor function of acute spinal cord injury rats. OECs transplantation combined with MP can more significantly promote recovery of the motor function of acute spinal cord injury rats. Related Articles | Metrics

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Repairing articular cartilage defects with cartilage-derived morphogenetic protein transfected autologous bone marrow mesenchymal stem cells Mu Chang-zheng, Ma Yun-sheng, Wang Zheng 2010, 14 (49):  9185.  doi: 10.3969/j.issn.1673-8225.2010.49.015 Abstract ( 378 )   PDF (620KB) ( 337 )   Save BACKGROUND: It can repairing cartilage defect somehow with scarffolds or stem cells, but we found that the effect of repair were not satisfied in large defects. The gene transfected stem cells can continuously release grow factors, and this may be presented a progress in cartilage defect. OBJECTIVE: To verify whether cartilage-derived morphogenetic protein 1 (CDMP1)-transfected autologous bone marrow mesenchymal stem cells (BMSCs) enhance cartilage repair in rabbit in vivo. METHODS: BMSCs were isolated from bone marrow of rabbits, and transfected with the CDMP1 gene by the lipofection method. The autologous cells were then implanted into full-thickness articular cartilage defects in the knee joints of rabbit as experimental group; BMSCs were transplanted as control BMSCs group and no cells transplanted as blank control group. The repair was analyzed by histological detection and histological score following surgery. RESULTS AND CONCLUSION: During in vivo repair of articular cartilage defects, cartilage regeneration was enhanced by the implantation of CDMP1 transfected autologous BMSCs. The defects were filled by hyaline cartilage and the deeper zone showed remodeling to subchondral bone overtime. There pair and reconstitution of zones of hyaline articular cartilage was superior to simple BMSCS implantation. The histological score of the CDMP1 transfected BMSCs group was significantly better than those of the BMSCs control group and the blank control group. These indicate that CDMP1 transfected BMSCs enhanced repair and remodeling of articular cartilage. Related Articles | Metrics

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Expression of recombinant coexpression plasmid of pIRES-hVEGF121cDNA/hBMP-4 in bone marrow mesenchymal stem cells and its effects on osteogenic differentiation Zhou Zhi-lin, Sun Jun, Zhao Yu, Wang Chun-lan 2010, 14 (49):  9189.  doi: 10.3969/j.issn.1673-8225.2010.49.016 Abstract ( 301 )   PDF (344KB) ( 314 )   Save BACKGROUND: Under some conditions, human body tissue repair is the result of common coordination of many factors. Thus, the construction of coexpression of many genetic carriers can meet the requirement of gene therapy of multiple factors, and is the new idea of recent gene therapy. OBJECTIVE: To observe the expression of recombinant coexpression plasmid of pIRES-hVEGF121cDNA/hBMP-4 in transfected rat bone marrow mesenchymal stem cells (BMSCs) and its effects on the differentiation of rat BMSCs into osteoblasts, and provide a basis for combination gene therapy of bone defects.  METHODS: BMSCs were isolated and cultivated from the bone marrow of rats by the whole bone marrow culture method. BMSCs were analyzed by the flow cytometry to detect the surface antigens. Recombinant coexpression plasmids of pIRES-hVEGF121cDNA/hBMP-4 were confirmed by restriction enzymolysis and then transfected rat BMSCs by lipofectamine. The total RNA and total proteins extracted from the resulting combination were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay. Alizarin red staining was utilized to detect the differentiation of rat BMSCs after transfection. RESULTS AND CONCLUSION: Flow cytometry analysis showed that BMSCs were strong positive for CD90, but did not express CD45. Recombinant coexpression plasmid of pIRES-hVEGF121cDNA/hBMP-4 was transfected successfully and the plasmid could coexpress hVEGF121cDNA and hBMP-4 efficiently in rat BMSCs. Alizarin red staining presented strong positive for 14 days after transfection. Human VEGF121 and BMP-4 gene can induce rat BMSCs to differentiate into osteoblasts, which provides a basis for combined gene therapy of bone defects. Related Articles | Metrics

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Adenoviral-mediated transfer of human transforming growth factor-beta 2 gene induces directional differentiation of bone marrow mesenchymal stem cells into chondrocytes Ju Xiao-dong, Ao Ying-fang, Wang Jian-quan, Deng Min 2010, 14 (49):  9194.  doi: 10.3969/j.issn.1673-8225.2010.49.017 Abstract ( 289 )   PDF (353KB) ( 431 )   Save BACKGROUND: Previous studies have shown that transforming growth factor-β1 (TGF-β1) contributes to proliferation of bone marrow mesenchymal stem cells (BMSCs), and induces the differentiation of BMSCs into chondrocytes. OBJECTIVE: To investigate the effect of human TGFβ2 (hTGFβ2) transfection on the directional differentiation of BMSCs into osteoblasts.   METHODS: A total of 16 clean male Lewis rats aged 3 weeks were selected for isolation of BMSCs. hTGFβ2 gene was transfected into the second passage of rat BMSCs cultured in vitro through Ad-hTGFβ2. 48 hours later, immunohistochemical staining, reverse transcription-polymerase chain reaction and Western blotting assay were used to determine target gene and cartilage specific protein-type Ⅱ collagen and aggrecan in transfected cells. RESULTS AND CONCLUSION: A population of BMSCs was successfully isolated from adult rats. They appeared fibroblast-likely and could stably proliferate and subculture in vitro. After transfected with Ad-hTGFβ2, the differentiation of bone marrow mesenchymal stem cells towards chondrocytes was verified by the positive results of type Ⅱ collagen protein expression through immunohistochemistry and Western blotting respectively. RT-PCR detected type Ⅱ collagen and aggrecan mRNA expression. Results suggest that MSCs from bone marrow presented multi-differentiational potential, and BMSCs can proliferate and subculture stably in vitro. Ad-hTGFβ2 is successfully transfected into BMSCs, and induces the differentiation of BMSCs into chondrocytes. Related Articles | Metrics

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Liposome-mediated pBLAST2-hHGF plasmid transfection in Lewis rat hepatic oval cells Li Zhu, Li Li, Ran Jiang-hua, Zhang Sheng-ning, Liu Jing, Liu Dian-sheng, Li Lai-bang, Chen Juan 2010, 14 (49):  9199.  doi: 10.3969/j.issn.1673-8225.2010.49.018 Abstract ( 376 )   PDF (304KB) ( 311 )   Save BACKGROUND: It is difficult to transfect exogenous gene into hepatocyte in vitro, while that is easily done in liver stem cells. The establishment of rat hepatic oval cells (HOCs) proliferation model is relatively stable at present, but stable culture and passage on primary HOCs are difficult. OBJECTIVE: To establish pBLAST2-hHGF plasmid stably transfected cell line in rat hepatic oval cells, and observe the biological characteristics of transfected cells. METHODS: Liposome-mediated gene transfection was used to transfer pBLAST2-hHGF plasmid into the fourth generation of hepatic oval cells derived from male Lewis rats. Morphological changes and cell proliferation of transfected cells were observed under inverted phase contrast microscope. Proliferation and differentiation capability of transfected cells were measured and stable culture conditions were explored. Western blot assay and enzyme linked immunosorbent assay were used to test human hepatocyte growth factor (hHGF) protein expression on transfected cells. RESULTS AND CONCLUSION: The fourth generation pBLAST2-hHGF/HOCs could be passaged steadily to 14 generations without significant morphological changes under the culture conditions containing various doses and types of cytokines. The growth and proliferation speed of transfected cells were obviously faster than that of HOCs. After removing growth factors mentioned above, pBLAST2-hHGF/HOCs quickly differentiated into hepatocytes and biliary epithelial cells in vitro. hHGF protein could be tested in pBLAST2-hHGF/HOCs using Western blot assay and it also could be tested with high level in culture fluid using enzyme linked immunosorbent assay. Results indicate that liposome-mediated gene transfection in this experiment was reliable and the stable passage time of pBLAST2-hHGF/HOC was longer than that of HOCs. pBLAST2-hHGF/HOCs had the ability to secrete hHGF protein and this cell line could be used in following experiment. Related Articles | Metrics

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Effects of JAK2-dependent ERK signal pathway on erythropoietin-induced functional activation of bone marrow-derived endothelial progenitor cells Zhao Wei, Yang Xiang-hong 2010, 14 (49):  9203.  doi: 10.3969/j.issn.1673-8225.2010.49.019 Abstract ( 290 )   PDF (394KB) ( 460 )   Save BACKGROUND: Erythropoietin (EPO) can promote the proliferation and migration of endothelial cells to form new vessels, so EPO may play an important role in functional activation of endothelial progenitor cells (EPCs). Except hematopoiesis, whether EPCs are a target of EPO remains poorly understood. OBJECTIVE: To investigate the effects of EPO on functional activation of EPCs and its signal pathway. METHODS: EPCs were separated from bone marrow of rats using density gradient centrifugation. First, cells were divided into control group (serum-free DMEM culture medium), recombinant human erythropoietin (rhEPO) in different dose groups (500,    1 000, 2 000 U/L), EPO (2 000 U/L) and the ERK inhibitor FR180204 (50 μmol/L) interaction group. Proliferation, migration, tubes formation and Bcl-2 expression of EPCs were detected with MTT assay, Transwell chamber, Matrigel and Western blot assay respectively. Secondly, cells were divided into control group (serum-free DMEM culture medium), rhEPO group (2 000 U/L), rhEPO (2 000 U/L) and JAK2 inhibitor SD1029 (10 µmol/L) interaction group. P-ERK expression of EPCs was detected by Western blot assay. RESULTS AND CONCLUSION: Compared with control group, EPO significantly increased EPCs proliferation, the number of migrating cells and tubes formation, up-regulated Bcl-2 protein expression in a dose-dependent manner in a certain rage. After added ERK inhibitor FR180204, these effects were significantly decreased compared with the rhEPO group (2 000 U/L). In addition, compared with the control group, the p-ERK1/2 expression in the rhEPO group (2000 U/L) was significantly up-regulated, JAK2 inhibitor SD1029 made the result reduced markedly. These indicate that EPO can promote functional activity of EPCs, which might be modulated by JAK2-dependent ERK signal pathway. Related Articles | Metrics

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Effects of dendritic cells-cytokine-induced killer cells against leukemia K562 cell lines in vitro Song Ying-ying, Su Rong-ying, Ai Li-mei 2010, 14 (49):  9208.  doi: 10.3969/j.issn.1673-8225.2010.49.020 Abstract ( 431 )   PDF (303KB) ( 503 )   Save BACKGROUND: The combined application of cytokine-induced killer cells (CIK) and dendritic cells to treatment of malignant tumors will help to remove T cell incompetence of some cancer patients, and plays synergistic anti-tumor effects. OBJECTIVE: To observe the effect of CIK cells co-cultured with dendritic cells against K562 cell lines. METHODS: CIK and dendritic cells were induced by peripheral blood mononuclear cells of normal persons. Dendritic cells and CIK were co-cultured for 3 days as effector cells. Immunophenotype of cells was measured by flow cytometry. K562 served as target cells, and peripheral blood mononuclear cells, CIK, dendritic cells, dendritic cells-CIK as effector cells. The killing activity was assayed by MTT assay. RESULTS AND CONCLUSION: With the increased ratio of effector to target, effector killing activity of each group also increased; for the same ratio, the killing activity of each group was different, of which dendritic cells-CIK was the strongest, up to (77.88± 1.57)% (P < 0.01). It also showed that dendritic cells-CIK has a stronger anti-tumor activity, and the toxicity of dendritic cells-CIK against leukemia cells was significantly higher than simple application of CIK or dendritic cells. Related Articles | Metrics

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Biological viability of rat bone marrow mesenchymal stem cells after the interference of astragalus mongholicus Sun Li, Jiao Bao-liang, Wang Jing-chuan, Wang Xin-sheng 2010, 14 (49):  9212.  doi: 10.3969/j.issn.1673-8225.2010.49.021 Abstract ( 237 )   PDF (243KB) ( 338 )   Save BACKGROUND: Astragalus mongholicus contributes to the differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons. OBJECTIVE: To observe effects of astragalus mongholicus on viability of rat BMSCs. METHODS: BMSCs of Sprague Dawley rats were isolated and cultured. The fourth passage of cells were treated with 50, 100, 200 g/L astragalus mongholicus for 5, 12, 24 hours respectively. BMSCs viability changes were observed by MTT assay after they were induced by astragalus mongholicus. RESULTS AND CONCLUSION: Primarily cultured cells were closed to 80%-90% fusion status after 8-10 days culture and they could be passaged. The third generation had been purified on the whole. The results from MTT assay showed that the BMSCs viability would be increased with the increased concentration of astragalus mongholicus and the prolonged action time. Morphological changes in neural cells appeared. There were significant differences between groups (P < 0.05). These indicate that astragalus mongholicus can induce the differentiation of BMSCs and nourish cells. Related Articles | Metrics

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Erythroid lineage differentiation of K562 cells induced by gensingnoside Re Lei Cui-rong, Jiang Rong, Wang Hong-ning, He Xuan, Zuo Guo-wei, Chen Di-long, Guan Tao, Wang Jian-wei 2010, 14 (49):  9216.  doi: 10.3969/j.issn.1673-8225.2010.49.022 Abstract ( 414 )   PDF (371KB) ( 344 )   Save BACKGROUND: Gensingnosides can promote normal hematopoiesis and inhibit proliferation of leukemia cells. Gensingnoside Re as the main active ingredient in Ginsenosides has a variety of activities. However, Re induces leukemia cells differentiation and the mechanisms remain unclear. OBJECTIVE: To investigate the effect of Gensingnoside Re on differentiation of chronic myelocytic leukemia K562 cells. METHODS: The density of K562 cells at logarithmic phase was regulated to 7×108/L. Cells in the blank control group were routinely cultured in RPMI1640 medium. Cells in the Gensingnoside Re group were incubated in RPMI1640 medium containing Gensingnoside Re with the final concentration of 30, 60, 90, 120, 150 μmol/L. Wright’s staining, benzidine staining and hemoglobinometry were utilized to determine the characteristics of K562 cell differentiation into erythroid cells. Expressions of markers on the surface of K562 cells were determined by flow cytometry.  RESULTS AND CONCLUSION: Gensingnoside Re 90 μmol/L could induce the K562 cell differentiation into erythroid cells. ① The morphological feature of K562 cells presented diminished volume, reduced nuclear diameter, abundant cytoplasm, and decreased karyoplasmic ratio. ②Hemoglobin in K562 cells induced by gensingnoside Re increased significantly in vitro. ③CD13 expressions on the surface of K562 cells increased. Related Articles | Metrics

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Promoting effect and mechanism of polygonatum medicated serum on bone marrow mesenchymal stem cells proliferation  Huang Jin, Zhang Jin, Xu Zhi-wei 2010, 14 (49):  9221.  doi: 10.3969/j.issn.1673-8225.2010.49.023 Abstract ( 308 )   PDF (282KB) ( 419 )   Save BACKGROUND: Current studies focus on the observation of single or complex Chinese medicine and serum containing drug on bone marrow mesenchymal stem cells (BMSCs) proliferation and differentiation, but few reports have addressed the mechanisms. OBJECTIVE: To investigate the effect of polygonatum medicated serum on BMSCs proliferation in vitro, and to analyze its action pathway and mechanism. METHODS: Male Sprague-Dawley rats were used for the isolation of BMSCs cells; the serum of female Sprague-Dawley rats was obtained after intragastric administration with medicine water solution. Rat BMSCs were isolated and purified by whole bone marrow adherent method, purified by trypsin. The third passage of BMSCs were prepared into single cell suspension, and incubated in the 96-well culture plate. The experimental group was added polygonatum medicated serum, while the control group added blank serum at concentration of 5%, 10%, 15%, 20%, 25%, 30%. The polygonatum medicated serum-induced proliferation effect on BMSCs was detected by MTT assay. Different cytokine mRNA expression was measured by RT-PCR. Granulocyte colony-stimulating factor mRNA expression was analyzed by real-time PCR. RESULTS AND CONCLUSION: Compare with the blank serum group, 10% polygonatum medicated serum significantly promoted the proliferation of BMSCs (P < 0.05), and significantly improved the expression of granulocyte colony-stimulating factor mRNA and protein (P < 0.05). Results suggest that 10% polygonatum medicated serum can promote proliferation of BMSCs, and the mechanism involves the stimulation of granulocyte colony-stimulating factor mRNA expression. Related Articles | Metrics

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Proliferative inhibition and differentiation of leukemia K562 cells induced by ginseng polysaccharide in vitro He Xuan, Jiang Rong, Li Jing, Zuo Guo-wei, Lei Cui-rong, Wang Ya-ping, Wang Jian-wei, Chen Di-long 2010, 14 (49):  9225.  doi: 10.3969/j.issn.1673-8225.2010.49.024 Abstract ( 363 )   PDF (417KB) ( 418 )   Save BACKGROUND: Ginseng polysaccharide can promote normal hematopoiesis and inhibit proliferation of leukemia K562 cells. However, their mechanisms have not been known. OBJECTIVE: To investigate the effect of ginseng polysaccharide on proliferation and differentiation of chronic myelocytic leukemia K562 cells in vitro. METHODS: K562 cells were kindly presented by Department of Clinical Laboratory, Chongqing Medical University. Ginseng polysaccharide was purchased from Shanxi Pude Medicines Co., Ltd. The density of K562 cells with logarithmic phase was regulated to 7×108/L. The conventional culture was performed in the blank control group. Ginseng polysaccharide group was incubated with 25, 50, 100, 200, 400, 600, 800 mg/L ginseng polysaccharide respectively. The proliferation of K562 cells was examined by MTT; Benzidine and Wright’s staining were used to observe the features of erythroid lineage differentiated from K562 cells. Meantime, the apoptotic rate of K562 cells was determined by flow cytometry. RESULTS AND CONCLUSION: Ginseng polysaccharide could significantly inhibit the proliferation and promote differentiation of K562 cells into erythroid linage cells in vitro. The proliferation of K562 cells was inhibited. The morphological feature of K562 cells tended to be more mature, showing small volume, reduced nuclear diameter, abundant cytoplasm, and decreased karyoplasmic ratio. Hemoglobin in K562 cells induced by ginseng polysaccharide increased significantly. Apoptotic rate of K562 cells treated with ginseng polysaccharide (100-800 mg/L) increased significantly in a concentration dependent fashion; the inhibitory rate reached a peak at 48 hours. These indicate that ginseng polysaccharide may inhibit the proliferation of K562 cells, promote their apoptosis and induce the differentiation of K562 cells into erythroid cells. Related Articles | Metrics

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Optimized isolation method and medium choice of embryonic stem cells from Kunming mice Tian Hai-qing, Cai Xia, La Xiao-lin, Hu Bo 2010, 14 (49):  9230.  doi: 10.3969/j.issn.1673-8225.2010.49.025 Abstract ( 308 )   PDF (403KB) ( 350 )   Save BACKGROUND: Up to date, hundreds of mouse embryonic stem cell (ESC) line has been established. Mouse of Kunming line is commonly used in experiment, and rare reports have addressed the success establishment. OBJECTIVE: To establish simple, efficient protocol for the derivation of ESCs from Kunming mice, and to elevate the success rate of establishment. METHODS: Mouse ESCs were incubated with the media containing serum surrogate or fetal bovine serum. Females of Kunming mice were superovulated with an injection of priganant mare’s serum gonadotropin and human chorionic gonadotropin. Blastocysts were collected by flushing the uterus at 3.5 days after copulation. Embryos were transferred in dish with fresh mouse embryonic fibroblasts feeder layer. Blastocysts detachment and colony forming efficiency of inner cell mass were observed. 4-6 days after culture, inner cell masses were digested mechanically in 0.05% trypsin-0.02%ethylenediamine tetraacetic acid.The disaggregated inner cell masses were transferred into a new feeder layer. Typical ESCs morphology was observed through an inverted microscope. RESULTS AND CONCLUSION: Pregnant mice of 3.5 days were the best choice to derive ESCs. Inner cell masses were separated through cutting in drops of 0.05% trypsin-0.02% ethylenediamine tetraacetic acid. The disaggregated inner cell masses grew more rapidly and maintained less differentiation. The ESCs were passaged many times in fetal bovine serum medium. This medium is more suitable to separate ESCs from Kunming mouse. Related Articles | Metrics

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Supportive effect of human amniotic mesenchymal stem cells on hematopoiesis in vitro Zhang Shou-qin, Li Wei-jia, Shi Ming-xia, Qian Yong-hai 2010, 14 (49):  9235.  doi: 10.3969/j.issn.1673-8225.2010.49.026 Abstract ( 197 )   PDF (298KB) ( 343 )   Save BACKGROUND: Bone marrow (BM) is the main source of mesenchymal stem cell (MSCs) at present, but because of the inconvenience of isolation, and the age-related decrease in the number of MSCs, amnion as a new source of MSCs has been widespreadly concerned recently. OBJECTIVE: To explore the supportive effects of MSCs derived from human amnions on hematopoiesis in vitro and to elevate the success rate of co-transplantation of amniotic mesenchymal stem cells (AMSCs) and hematopoietic stem cells. METHODS: MSCs were isolated and cultured from human amnions after childbirth by the tissue pieces culture method. Mononuclear cells (MNCs) were isolated from umbilical cord blood using density gradient centrifugation. CD34+ cells were isolated by magnetic cell sorting. The MNC and CD34+ cells were co-cultured with AMSCs for four weeks respectively. The concentration of the floating cells was recorded weekly. And as the control, they were co-cultured with the BM-MSCs and deeder layer-free group respectively as well. The MNC and CD34-positive cells were plated in semisolid culture medium separately, and the colony formation was assessed at 14 days after the plating. RESULTS AND CONCLUSION: AMSCs promoted the expansion of human cord blood MNCs and CD34+ cells, and also promoted the formation of hematopoietic progenitor cell colony in methylcellulose semisolid culture medium. Its capacity of hematopoitic support was similar to that of BM-MSCs, without significant difference. These indicate that in vitro AMSCs had the capacity for hematopoietic support similar to BM-MSCs. Thus, they may be an ideal source of stem cells for the expansion of hematopoietic stem cells in vitro and the co-transplantation of MSCs and hematopoietic stem cells to improve the success rate of clinical transplantations. Related Articles | Metrics

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Differentiation of modified method cultured human dental pulp mesenchymal stem cells into neuron-like cells Li Yuan, Guan Fang-xia, Li Ji, Yang Bo, Hu Xiang, Du Ying, Gu Chen-xi, Lei Ning-jing, Wang Xiao-wei 2010, 14 (49):  9239.  doi: 10.3969/j.issn.1673-8225.2010.49.027 Abstract ( 249 )   PDF (399KB) ( 355 )   Save BACKGROUND: The success rate of dental pulp stem cells (DPSCs) which isolated by the modified cultured method of tissue is elevated. However, few reports have concerned the difference in biological features between DPSCs and stem cells extracted by emzyme digestion method. OBJECTIVE: To observe the difference in the differentiation of DPSCs isolated by the new method and isolated by traditional method into neuron-like cells. METHODS: The specimen of the same tissue was randomly divided into two parts: one is isolated by the modified cultured method of tissue (A group), another is isolated by the traditional method (B group). Growth, antigen expression and differentiation of DPSCs into neuron-like cells were observed and compared using immunohistochemical method, flow cytometry and reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: There were no distinct changes in morphology expression, antigen ability, differentiation ability into neurons between A group and B group. However, A group had simple process and obtained a large quantity. Related Articles | Metrics

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Application of rabbit adipose-derived stem cells to the expanded flap with congestion Liang Jiu-long, Liu Xiao-yan, Tao Kai, Li Xia 2010, 14 (49):  9244.  doi: 10.3969/j.issn.1673-8225.2010.49.028 Abstract ( 304 )   PDF (350KB) ( 434 )   Save BACKGROUND: To expand skin flap congestion is a difficulty in the plastic surgery. Previous methods are limited in application. Adipose-derived stem cells (ADSCs) can prolong the length of survived expanded skin flap. OBJECTIVE: To solve the problem of congestion in the expanded flap by rabbit ADSCs. METHODS: A total of 20 New Zealand rabbits were used to establish models of expanded flap congestion. 10 mL expandor was implanted in the skin of the back of the ear. Water was regularly injected. On day 20, the expandor was obtained to form expanded congestion flap. Autologous ADSCs were injected into one side of the congestion flap as experimental group. Saline was injected into another side as control group. At 2 weeks following injection, the survival of the flap was observed. Pathological sections were observed. Immunohistochemistry staining for CD34 was performed to quantitatively analyze vascularization. RESULTS AND CONCLUSION: The survived length of flap was significantly longer in the experimental group than the control group (P < 0.01). Pathological sections demonstrated that the density of CD34 staining was significantly greater in the experimental group compared with the control group (P < 0.01). Results indicated that ADSCs can increase vascularization, improve congestion of the flap, and elevate the length in the expanded flap. Related Articles | Metrics

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Changes in the number of endothelial progenitor cells from peripheral blood in patients with coronary atherosclerotic heart disease before and after extracorporeal cardiac shock wave therapy Cai Hong-yan, Wang Yu, Li Lin, Zhao Ling, Xiao Jian-ming, Cao Jing-ming, Yang Xi-yun, Yang Chao, Guo Tao 2010, 14 (49):  9249.  doi: 10.3969/j.issn.1673-8225.2010.49.029 Abstract ( 253 )   PDF (330KB) ( 411 )   Save BACKGROUND: The mechanism underlying the effect of extracorporeal cardiac shock wave therapy on endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary atherosclerotic heart disease (CAHD) is poorly understood. OBJECTIVE: To observe the effects of cardiac shock wave therapy on the proliferation and the differentiation of EPCs from peripheral blood in CAHD patients. METHODS: A total of 25 CAHD patients undergoing extracorporeal cardiac shock wave therapy were selected, and another 10 patients who did not receive extracorporeal cardiac shock wave therapy served as controls. Mononuclear cells obtained from peripheral blood were cultured in EGM-2-MV medium. Cell morphology and the number of colonies formed were observed and analyzed. After 7 days of culture, adherent cells were analyzed and counted. Whether EPCs differentiated or not was identified by laser confocal microscopy. RESULTS AND CONCLUSION: The number of EPCs from peripheral blood and colonies formed were significantly increased (P < 0.05) compared with the control group. It is indicated that extracorporeal cardiac shock wave therapy can promote the proliferation and differentiation of EPCs from peripheral blood into endothelial cell lineage in CAHD patients. Related Articles | Metrics

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Apoptosis of lung mesenchymal cells under normal oxygen culture versus low oxygen culture Xing Wen, Yang Shao-guang, Han Zhi-bo, Ru Yong-xin, Liu Meng, Lu Shi-hong, Zhao Qin-jun, Li Jian-ping, Ren Qian, Han Zhong-chao 2010, 14 (49):  9253.  doi: 10.3969/j.issn.1673-8225.2010.49.030 Abstract ( 406 )   PDF (409KB) ( 347 )   Save BACKGROUND: Expansion of murine-derived mesenchymal cells is difficult in vitro and it is unclear whether this phenomenon is caused partly by cell apoptosis. OBJECTIVE: To compare the differences of mouse lung mesenchymal cell apoptosis between normal oxygen and low oxygen culture conditions. METHODS: C57BL/6J male mouse lung derived-mesenchymal cells were isolated by primary explant culture and cultured under normal (20% O2) and low oxygen (5% O2) conditions. The differences in intracellular active oxygen level, apoptotic rate and ultrastructure were observed. RESULTS AND CONCLUSION: The number of hematopoietic cells was decreased with increased cell passage under both 5% and 20% O2 condition. The active oxygen levels were significantly greater in the mouse lung-mesenchymal cells under 20% O2 condition compared with those under 5% O2 condition. Apoptosis was more in cells under 20% O2 condition compared with those under 5% O2 condition. These indicate that under normal oxygen culture condition, the apoptotic rate of cells increases and low oxygen can reduce the apoptosis. Related Articles | Metrics

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Differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells in vitro Chen Jun, Liu Yu-xia, Jiang Wen-hua, Han Guang, Jiang Wen-ping 2010, 14 (49):  9257.  doi: 10.3969/j.issn.1673-8225.2010.49.031 Abstract ( 250 )   PDF (1101KB) ( 353 )   Save BACKGROUND: Many studies have shown that mesenchymal stem cells have the potential to differentiate into hepatocytes, which will not only provide seed cell source for hepatocellular transplantation, bioartificial liver and hepatic tissue engineering, but also open breakthrough therapies in acute liver failure, end-stage liver disease and genetic metabolic liver disease. OBJECTIVE: To induce human umbilical cord mesenchymal stem cells into hepatocyte-like cells in vitro, to observe morphological and functional changes respectively. METHODS: The logarithmic phase of human umbilical cord mesenchymal stem cells of P3 was cultured at the density of 1×109/L in culture flasks containing DMEM/F12 with 10% fetal bovine serum (FBS). The cells were divided into 3 groups: group 1 treated with DMEM/F12 medium, containing 10% FBS, 20 μg/L hepatocyte growth factor (HGF), 10 μg/L basic fibroblast growth factor (bFGF), group 2 treated with DMEM/F12 medium, containing 10% FBS, 20 μg/L HGF, 10 μg/L bFGF, 20 μg/L oncostatin, blank control group treated without growth factor. The medium was changed 2-3 times a week according to cell growth. The cell morphology was observed at 7, 14, 18 days following culture under an inverted microscope. Expressions of alpha fetoprotein, albumin and cytokeratin18 were examined by immunocytochemistry. The glycogen deposits were examined by periodic acid-Schiff method. RESULTS AND CONCLUSION: The human umbilical cord mesenchymal stem cells could be differentiated into heaptocyte-like cells by DMEM/F12 medium supplemented with 10% FBS, 20 μg/L HGF, 10 μg/L bFGF and 20 μg/L oncostatin. The induced efficiency was more than by DMEM/F12 medium supplemented with 10% FBS, 20 μg/L HGF, 10 μg/L bFGF. Related Articles | Metrics

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Application of bone marrow mesenchymal stem cells to liver failure Yang Ling-ling, He Jin-qiu 2010, 14 (49):  9263.  doi: 10.3969/j.issn.1673-8225.2010.49.034 Abstract ( 304 )   PDF (264KB) ( 264 )   Save BACKGROUND: Recent studies have shown that bone marrow mesenchymal stem cells (BMSCs) could differentiate into hepatocyte-like cells in pathologic liver tissue and restore the injured livers.  OBJECTIVE: To conclude and analyze biological characteristics of BMSCs and their clinical application in liver failure treatment.  METHODS: We retrieved China National Knowledge Infrastructure (CNKI) and PubMed databases for articles concerning research and application of BMSCs in liver failure treatment published from January 1997 to December 2009. The key words were “bone marrow mesenchymal stem cells, liver failure” or “liver stem cells, liver disease”. The articles addressing BMSCs for liver failure, and the articles published recently or in authorized journals, were included. A total of 371 literatures were primarily selected, and 31 were included in accordance with inclusion criteria. RESULTS AND CONCLUSION: BMSCs are a type of adult stem cells, which possess an extensive self-renewal potential and ability to differentiate into various cell types. Their ability of repairing the liver failure is by the mechanisms of transdifferentiation and cell fusion. With the development of research, BMSCs transplantation is gradually applied to treatment on liver failure. However, the study deserves further improvement. Related Articles | Metrics

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Effects of bone marrow mesenchymal stem cells on transplantation immunity Liu Yin-mei, Zhang Peng 2010, 14 (49):  9264.  doi: 10.3969/j.issn.1673-8225.2010.49.033 Abstract ( 263 )   PDF (303KB) ( 378 )   Save BACKGROUND: Recent studies have demonstrated that bone marrow mesenchymal stem cells (BMSCs) can support in vitro hematopoiesis, promote reconstruction of in vivo hematopoietic function, have low immunogenicity and inhibit proliferation of allogenic T lymphocytes, and play important roles in regulating transplantation immunity. However, its mechanism remains unclear. OBJECTIVE: To summarize the immunoloregulation characteristics of BMSCs and its effects on hematopoietic stem cell transplantation. METHODS: The first author searched PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) published from 2000 to 2008 for articles on effects of BMSCs on transplantation immunity. The key words were “BMSCs, hematopoietic stem cell transplantation, transplantation immunity”. Repetitive studies were excluded. Finally, 37 articles were included for this review. RESULTS AND CONCLUSION: BMSCs are stem cells in the bone marrow besides hematopoietic stem cells. In immune reaction, BMSCs can avoid immunological recognition, and suppress immune reaction. The immunesuppressive effects of BMSCs are effectively used in hematopoietic stem cell transplantation, can decrease graft versus host disease, promote graft survival, and have wide application prospect. However, at present, there is no unified standard to identify BMSCs, and the specific marker of BMSCs has not been screened. Some problems should be further investigated such as lack of safety assessment, whether BMSCs affected graft versus leukemia when affected graft versus host disease, as well as the effects on tumor cell growth. Related Articles | Metrics

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Osteogenic differentiation and clinical application of bone marrow mesenchymal stem cells Chen Guang-dong, Yang Hui-lin, Wang Gen-lin, Wang Li-jun 2010, 14 (49):  9272.  doi: 10.3969/j.issn.1673-8225.2010.49.035 Abstract ( 397 )   PDF (348KB) ( 425 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have potential to differentiate into osteoblasts and have great prospect in application as new source of seed cells in tissue engineering. However, there are still some problems such as isolation and purification of BMSCs, optimal density for differentiation into osteoblasts, oncogenicity and safety in vivo, deserving more attention and further investigation. OBJECTIVE: Basic research and clinical application of BMSCs are extensive. There are still lots of problems deserving more concern. This study reviewed characteristics of biology, the induced differentiation, isolation and identification of BMSCs, and clinical applicability of BMSCs. METHODS: We retrieved the articles in Chinese Journal Full-Text Database, PubMed database and Ovid database published from 1997 to 2010. Key words were “mesenchymal stem cells, osteoblast cells”. Articles concerning BMSCs were included, and literatures of repetitive content and lack of originality were excluded. A total of 31 literatures were obtained for further analysis. RESULTS AND CONCLUSION: BMSCs were isolated from bone marrow mainly using density gradient centrifugation, immunology separation and adherence screening method at present. The main technical characteristics of differentiation of BMSCs into osteoblasts are to transfer gene with osteogenic effect into target cells; transcription into mRNA using target cells and translation into protein with osteogenic effects in the lesion site; oneself ossification was promoted through persistent effects of target cells. BMSCs depended on biological stents can rapidly differentiate into osteoblasts and proliferate under suitable induction of growth factors. Patients who suffered from fracture nonunion and metabolic bone diseases have benefited from clinical application of BMSCs. Related Articles | Metrics

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Bone marrow mesenchymal stem cells and sports knee injury Zhong Dong-hong, Chen Hui-yu 2010, 14 (49):  9277.  doi: 10.3969/j.issn.1673-8225.2010.49.036 Abstract ( 300 )   PDF (282KB) ( 307 )   Save BACKGROUND: Knee injury is one of the most common sports injuries, and it seriously impacts the athletes training and competition. The injury can also cause cardiovascular and metabolic adaptation to decline and disuse muscle atrophy. Bone marrow mesenchymal stem cells (BMSCs) characterized by pluripotent, low immunogenicity, no rejection of its advantages provide a broad application prospects in prevention and treatment of knee joint injury. OBJECTIVE: To summarize the application of BMSCs to sports knee injury, and to provide a scientific basis for prevention and treatment of knee injury. METHODS: We searched PubMed database for literatures published from January 1994 to May 2010. Key words were “bone marrow mesenchymal stem cells, exercise, knee injury”. Articles on BMSCs and exercise knee injury were included, and repetitive studies were excluded. Finally, 34 articles were selected for further analysis. RESULTS AND CONCLUSION: The repair effect of BMSCs on knee joint cartilage, tendon, muscle cells and meniscus was explored by analyzing physiological characteristics, clinical application of BMSCs and causes of sports knee injury. The results showed that BMSCs for its convenience, less invasive and easy separation from the bone marrow have great values for repairing knee cartilage damage, promoting tendon repair and treating knee joint injury. Related Articles | Metrics

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Application of induced differentiation of bone marrow mesenchymal stem cells to retinal diseases Zhang Ke-jian, Liu Xue-zheng, Hou Yang,Zhang Ming-rui 2010, 14 (49):  9281.  doi: 10.3969/j.issn.1673-8225.2010.037 Abstract ( 373 )   PDF (339KB) ( 405 )   Save BACKGROUND: Recent studies indicate that bone marrow mesenchymal stem cells (BMSCs) could differentiate into neuron-like cells, have the fusion with the original retina structure and can be part of the treatment of retinal degeneration.  OBJECTIVE: To conclude and analyze biological characteristics of BMSCs and their clinical application in retinopathy.  METHODS: China National Knowledge Infrastructure and PubMed Database were searched by computer for articles on research and application of BMSCs in retinopathy published from January 1995 to May 2010. The key words were “bone marrow mesenchymal stem cells, retina” or “retinopathy”. The related articles about BMSCs for retinopathy were included, and those published recently or in authorized journals in the same field were included. A total of 286 articles were primarily collected. In accordance with inclusion criteria, 29 articles were used for this review. RESULTS AND CONCLUSION: BMSCs have the potential of differentiating into retina-like cells, and possibility of differentiating into retinal nerve cells to repair damaged retina. BMSCs can differentiate into retinal nerve cells, help damaged retina to repair. BMSCs transplantation in retina can be used to treat retinal degeneration. However, at present, it is not ideal to differentiate into retinal cells following BMSCs transplantation. The mechanism of induced differentiation remains unclear. Thus, the pathway of retinal transplantation and posttransplanted outcomes require further investigations. Related Articles | Metrics

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Traditional Chinese medicine in stem cells study Zhang Liang-deng Yao Kui-wu, Wang Jie 2010, 14 (49):  9285.  doi: 10.3969/j.issn.1673-8225.2010.49.038 Abstract ( 315 )   PDF (291KB) ( 433 )   Save BACKGROUND: Stem cells are hot topic research in regenerative medicine. More and more literatures on traditional Chinese medicine (TCM) intervention in stem cell culture have been reported. OBJECTIVE: To further explore the correlation of TCM to culture, induction, growth and differentiation of stem cells by reviewing and summarizing studies of stem cells intervention in TCM recent years. METHODS: We searched experiments or clinical researches literatures of stem cells intervention in TCM through China National Knowledge Infrastructure and Chinese sci-tech periodical full-text database published from January 1991 to January 2010. The key words were “TCM” or “Chinese materia medica” and second key word “stem cells”. Then, TCM prescriptions, stem cells types, application value, affiliations and funds were extracted, for setting up a database and frequency analysis.  RESULTS AND CONCLUSION: Totally 167 documents were retrieved, only 31 of which met the inclusion criteria. The usual therapies on stem cells intervention in TCM are that removing stasis and blood circulation, tonifying qi, nourishing kidney and replenish vital essence. There is regularity between TCM therapies and stem cells types. Famous and effective prescriptions and effective TCM components are widely used, especially TCM of notifying qi, removing stasis and blood circulation, nourishing kidney and replenish vital essence. TCM can improve mobilization, proliferation and differentiation of stem cells to repair injured tissues and organs. Fund increased year by year, mainly focused by National Nature Science Foundation of China and related researches have focused on developed regions. There is solid base of theory and clinic of targeted stem cells in TCM. Targeted stem cells in TCM would integrate regenerative thought with clinical thought of TCM, extend the harmony of yin and yang to the era of stem cells and regenerative medicine. Related Articles | Metrics

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Biological characteristics and application of adipose tissue-derived stem cells in tissue engineering  Wei Mao-ping, Liu Yu-mei, Zhang Zi-qiang 2010, 14 (49):  9290.  doi: 10.3969/j.issn.1673-8225.2010.49.039 Abstract ( 252 )   PDF (380KB) ( 290 )   Save BACKGROUND: Adipose tissue-derived stem cells possess self-renewal and multi-directional differentiation potential. In vitro it could be induced to different cells in specific differentiation medium, such as lipocytes, skeltogenous cells, cartilage cells, cardiac muscle cells, nerve cells and liver cells. These characteristics show that adipose tissue-derived stem cells possess favorable application prospect.  OBJECTIVE: To understand the biological characteristics of adipose-derived mesenchymal stem cells, and to explore its clinical application in tissue engineering field. METHODS: The databases of Elsevier (2000-01/2010-05) and China National Knowledge Infrastructure (2000-01/2010-05) were searched with key words of “adipose tissue-derived stem cell, tissue engineering”. Literature search was limited to English and Chinese languages. RESULTS AND CONCLUSION: Adipose-derived stem cells could proliferate rapidly, and material was convenient to harvest. It could be obtained by itself, and also avoid immunological rejection. At present, there was no gold standard to identify adipose-derived stem cells, but investigators had observed in vitro cultured adipose-derived stem cells that had the specific surface labeling of mesenchymal stem cells using flow cytometry and immunohistochemical method. Adipose-derived stem cells could differentiate into adipocytes, osteocytes, chondrocytes, muscle cells, hematopoietic cells, liver cells and nerve cells. Tissue or organ defect disease, degenerative disease and hereditary disease can use tissue engineering technique to implant tissue-derived stem cells and stents into the bodies, which has been a research focus in clinic. Related Articles | Metrics

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Stem cell application in spinal cord injury: Transplantation time, method, frequency and cell selection  Li Ya-mei, Jia Gong-wei, Yu Le-hua 2010, 14 (49):  9294.  doi: 10.3969/j.issn.1673-8225.2010.49.040 Abstract ( 239 )   PDF (260KB) ( 393 )   Save BACKGROUND: Tissue regeneration and functional recovery after spinal cord injury (SCI) have always been a significant challenge for neurosurgeon and rehabilitation physicians. For decades, a lot of animal experiments have confirmed that stem cell transplantation had certain repair effects on SCI, and had made many clinical trials, but did not obtain satisfactory effects. OBJECTIVE: To review recent advances and problems in stem cell transplantation for SCI, so as to point the way for further studies, and to promote recovery of SCI patients. METHODS: We retrieved articles about therapy of stem cell for SCI in Vip, Wanfang and PubMed databases for articles published from January 1990 to March 2010. The key words were “spinal cord injury, stem cells, transplantation, mechanism, clinical application” for retrieval in the title and abstracts. A total of 285 articles were primarily obtained, and 30 articles were included for this review in accordance with inclusion criteria and exclusion criteria. RESULTS AND CONCLUSION: According to the inclusion and exclusion criteria, we got 30 articles. Through reading and collecting information, we summarized the basic mechanisms of stem cell transplantation for treating SCI, sources and selection of stem cells, transplantation methods, transplantation frequency and the clinical status, and put forward the problems at present. Stem cell transplantation for SCI is a great potential treatment, but some problems also exist. These problems will be the direction of further research addressing stem cell therapy for SCI. Related Articles | Metrics

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Effects of miRNA and Notch signaling pathway on cardiac development Huang Feng, Zhou Sheng-hua, Pan Jia-yi, Bai Zhong-le, Wang Zhen-he 2010, 14 (49):  9298.  doi: 10.3969/j.issn.1673-8225.2010.49.041 Abstract ( 351 )   PDF (324KB) ( 438 )   Save BACKGROUND: miRNA has the functions of regulating cell proliferation and differentiation, promoting the development of organisms. Notch signaling pathway plays an important role in cell proliferation and differentiation, and the same in organ and organization development. Studies concerning miRNA and Notch signaling pathways in cardiac development and their relationship may provide new ideas in stem cell transplantation for treating certain heart diseases. OBJECTIVE: To summarize the study progress of the roles that miRNA and Notch signaling pathway playing in cardiac development. METHODS: We searched literary on the Google Scholar, PubMed database and ScienceDirect-Home website with the key words of “miRNA, Notch signaling pathway, cardiac development, stem cells”. RESULTS AND CONCLUSION: miRNA has many biological functions in cardiac development such as regulation of cell proliferation and differentiation by adjusting target gene mRNA, or inhibiting its translation. Notch signaling pathway could determine the fate of cells and play an important role in cardiac normal development. Moreover, miRNA could induce embryonic stem cells differentiation into myocardial cells by regulating Dll1 receptor of the Notch signal. Grasping the mechanism and relation of miRNA or Notch signaling pathway playing in cardiac development, and how they induce stem cells differentiation into myocardial cells, could provide us with new intervention targets using stem cells to treat certain heart diseases. Related Articles | Metrics

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Effects of natural killer cells on graft rejection, hematopoietic and immune reconstitution following allogeneic bone marrow transplantation Yang Zhi-gang, Xiong Dan, Zeng Yao-ying, Liang Liang, Li Qing-hua, Wu Guo-cai 2010, 14 (49):  9303.  doi: 10.3969/j.issn.1673-8225.2010.49.042 Abstract ( 335 )   PDF (240KB) ( 396 )   Save BACKGROUND: The connection between Natural killer (NK)-cells and allogeneic bone marrow transplantation (allo-BMT) has aroused increasing attention. OBJECTIVE: To explore the effect of NK cells on graft rejection, hematopoietic and immune reconstitution in mouse undergoing allo-BMT. METHODS: Lethally and nonlethally irradiated BALB/c (H-2d) mice were transplanted with C57BL/6(H-2b) bone marrow plus donor peripheral T cells and/or NK cells. RESULTS AND CONCLUSION: Compared with lethally irradiated and allo-BMT group without infusion of NK cells, the survival rate in lethally irradiated and allo-BMT group with infusion of NK cells significantly enhanced; leukocytes count, expression level of CD19+ and CD34+ cell count recovered rapidly; expression level of H-2b+ cell obviously increased. Expression level of CD34+ cell in the group with infusion of NK cells was obviously lower than that of the group without infusion of NK cells at 28 days after transplantation, but there was no significant difference between the 2 groups at 60 days (P > 0.05). In nonlethally irradiated and allo-BMT group without NK cell infusion, expression level of H-2b+ cell significantly decreased at 30 days after transplantation, and reduced to before transplantation level at 60 days; while expression of H-2b+ cell yet could be detected with more than 80% at 60 days after transplantation in group infused with high and low concentration of NK cells. In allo-BMT mice, alloreactive NK cell inhibits graft rejection, enhances engraftment, promotes the reconstitution of hematopoiesis and immunity, and increases survival rates. Related Articles | Metrics

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Induction of umbilical cord blood-derived mesenchymal stem cells differentiating into pancreatic islet β-like cells in vitro Chi Zuo-hua, Lu Yan, Zhang Yuan 2010, 14 (49):  9307.  doi: 10.3969/j.issn.1673-8225.2010.49.043 Abstract ( 200 )   PDF (340KB) ( 358 )   Save BACKGROUND: At present, there are many reports regarding the differentiation of bone marrow-derived mesenchymal stem cells or pancreatic gland stem cells into pancreatic islet β-like cells. But little is about umbilical cord blood-derived mesenchymal stem cells (UCBMSCs) differentiation into pancreatic islet β-like cells in vitro. OBJECTIVE: To investigate whether UCBMSCs can differentiate into pancreatic islet β-like cells in vitro and the optimal inducing condition. METHODS: UCB samples were obtained sterilely from healthy parturients. Nucleated cells were isolated by sedimentation with hydroxyethyl starch and MSCs were obtained by adherent method. Then purified UCBMSCs were induced with epidermal growth factor, β-mercaptoethanol, high glucose, Activin A and hepatocyte growth factor (HGF). Following cell morphology observation, induced cells were identified by insulin immunofluorescence. In addition, insulin secretion and glucose-stimulated insulin release were examined by chemiluminescence immunoassay.    RESULTS AND CONCLUSION: After induction, many cells exhibited a round appearance and produced islet-like cell clusters. Immunofluorescence assay showed insulin positive in the treated cells. In addition, chemiluminescence immunoassay demonstrated low expression of insulin and secretion of insulin upon glucose challenge. UCBMSCs can differentiate into pancreatic islet β-like cells in vitro.  Related Articles | Metrics