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03 September 2013, Volume 17 Issue 36 Previous Issue    Next Issue

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Green fluorescent protein expression in vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cells Yang Yang, Huang Zheng-de, Tian Xue-fei, Zhou De-sheng, Li Xin-hui, Hu Hua, Yang Yan-hong 2013, 17 (36):  6381-6387.  doi: 10.3969/j.issn.2095-4344.2013.36.001 Abstract ( 292 )   PDF (1914KB) ( 404 )   Save BACKGROUND: Vascular endothelial growth factor is a potent angiogenesis and permeability inducible factor. Vascular endothelial growth factor 165 and vascular endothelial growth factor 121 are mainly expressed in vivo, with a strong role of angiogenesis. OBJECTIVE: To observe the feasibility of vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cells to differentiate into vascular endothelial cells. METHODS: Bone marrow derived mesenchymal stem cells were isolated and collected from 50 g Sprague-Dawley rats,and identified by flow cytometry. The plasmid pGLV-EF1a carrying a vascular endothelial growth factor 165 gene was transfected to the mesenchymal stem cells using lentiviral. Expression of green fluorescent protein was observed under a fluorescence microscope. RESULTS AND CONCLUSION: After 12 hours of transfection, expression of green fluorescent protein was observed, increased at 48 hours, peaked at 72 hours and gradually declined thereafter. Results prove that vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cells have the expression of green fluorescent protein, indicating successful transfection. It is feasible to induce bone marrow mesenchymal stem cells to differentiate into vascular endothelial cells. Figures and Tables | References | Related Articles | Metrics

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Correlation between magnitude and duration of hydrostatic pressure and the differentiation of human bone marrow mesenchymal stem cells He Chuan, Liang Jing, Deng Lian-fu, Feng Jian-min 2013, 17 (36):  6388-6395.  doi: 10.3969/j.issn.2095-4344.2013.36.002 Abstract ( 235 )   PDF (520KB) ( 304 )   Save BACKGROUND: Mechanical signal has close correlation with the growth, development, repair and reconstruction of the skeletal system and the development of disease, the effect and the mechanism on bone marrow mesenchymal stem cells is worthy to concern.  OBJECTIVE: To explore the effect and mechanism of hydrostatic pressures on the differentiation of bone marrow mesenchymal stem cells.  METHODS: Short-term experiment: the human bone marrow mesenchymal stem cells were incubated into the normal Dulbecco’s modified Eagle’s medium, osteogenic medium or the Dulbecco’s modified Eagle’s medium containing extracellular signal-regulated kinase 1/2 inhibitor U0126, respectively. Homemade pressure loading system was used to impose 0, 40 and 80 kPa hydrostatic pressure for 1 and 4 hours. Long-term experiment: human bone marrow mesenchymal stem cells were incubated into the normal Dulbecco’s modified Eagle’s medium or osteogenic medium respectively, and then 40 kPa hydrostatic pressures was loaded for 4 hours per day, and lasted for 14 days. The cells without hydrostatic pressure were regarded as the control group. RESULTS AND CONCLUSION: Real-time quantitative reverse transcription PCR results showed that after osteogenic induction and simulated with 40 kPa hydrostatic pressure for 4 hours, the mRNA expressions of core binding factor α1 and osteocalcin in the bone marrow mesenchymal stem cells were increased, while the mRNA expressions of peroxisome proliferator-activated receptor γ2 and adipsin were decreased, and the 80 kPa hydrostatic pressure did not cause such reactivity. The osteogenic induction effect of 40 kPa hydrostatic pressure could be partial antagonized with U0126. Histochemical staining showed that after simulated with 40 kPa hydrostatic pressure for 7 days, the expression and activity of alkaline phosphatase of bone marrow mesenchymal stem cells were increased; after lasted for 14 days, the mRNA expressions of peroxisome proliferator-activated receptor γ2 and adipsin were increased. Certain intensity and duration of hydrostatic pressure stimulation can regulate the differentiation of bone marrow mesenchymal stem cells, and the mechanism is only partly mediated by the extracellular signal-regulated kinase 1/2 signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Proliferation and differentiation of rat bone marrow mesenchymal stem cells under different mechanical strains Wang Qiu-shi, Yang Xiao-qin, Zhu Xiao-wen, Hu Jing, Zou Shu-juan 2013, 17 (36):  6396-6402.  doi: 10.3969/j.issn.2095-4344.2013.36.003 Abstract ( 284 )   PDF (868KB) ( 427 )   Save BACKGROUND: In vitro and in vivo studies of cell response to a variety of mechanical loadings have demonstrated the stimulation of bone formation by loads. However, the effects of different mechanical strains on the same cells have never been adequately studied by far. OBJECTIVE: To investigate the effects of different mechanical strains on rat bone marrow mesenchymal stem cells. METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro. Bone marrow mesenchymal stem cells were subjected to different stimulations including dynamic stretch, static stretch and hybrid stretch through the use of custom-made mechanical stretch device. Cellular proliferation, alkaline phosphatase activity and mRNA expression of Runx2 of bone marrow mesenchymal stem cells were detected and the secretion of osteocalcin was evaluated under three different stretch modes respectively. RESULTS AND CONCLUSION: Compared to the control group, cell proliferation increased by 18.67%, however, alkaline phosphatase activity, Runx2 expression and osteocalcin secretion were not changed obviously in the static stretchgroup. Compared to the control group, alkaline phosphatase activity, Runx2 expression and osteocalcin secretion increased by 60.33%, 49.67% and 48% respectively; however, cell proliferation was inhibited, in the dynamic stretch group. Compared to the control group, cell proliferation was slightly, but not significantly, increased in the hybrid stretch group, and the alkaline phosphatase activity, Runx2 expression and osteocalcin secretion increased although the increases were not as apparent as those in the dynamic stretch group. These findings suggest that static mechanical strain can significantly promote cell proliferation, the dynamic mechanical strain more greatly promotes osteogenic differentiation of bone marrow mesenchymal stem cells, and the hybrid mechanical strain promotes the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. Figures and Tables | References | Related Articles | Metrics

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Restraining degeneration of intervertebral discs by transplantation of Notch-1knockout rabbit bone marrow mesenchymal stem cells Mori Gele, Shao Zeng-wu 2013, 17 (36):  6403-6408.  doi: 10.3969/j.issn.2095-4344.2013.36.004 Abstract ( 464 )   PDF (2267KB) ( 367 )   Save BACKGROUND: Preliminary experiments have demonstrated that the expression of Notch-1 in the degenerated intervertebral disc was increased. However, the role of Notch-1 in the nucleus pulposus-like cell differentiation of mesenchymal stem cells remains unclear. OBJECTIVE: To investigate the bone marrow mesenchymal stem cells in suppressing the degeneration of intervertebral discs after Notch1 gene knockout. METHODS: (1) Bone marrow mesenchymal stem cells were isolated from the femur bone of four New Zealand rabbits weighing 0.4-0.5 kg, under deep anesthesia, and then purified with discontinuous gradient density centrifugation method. (2) Notch1 shRNAs and blank plasmid shRNA were designed, synthesized, and transiently transfected into these mesenchymal stem cells, and the differentiation of mesenchymal stem cells was induced with transforming growth factor beta-1. (3) Ten New Zealand rabbits weighing 1.0-1.5 kg were involved in this study. The rabbits’ intervertebral discs in L3-4, L4-5 and L5-6 were stabbed by a needle, and nucleus pulposus tissue was harvested for modeling. The cells were divided into blank plasmid transfected with transforming growth factor beta-1 group, shRNA-Notch-1 plasmid transfected with transforming growth factor beta-1 group, and non-transfected treated with transforming growth factor beta-1 group. Two weeks later the treated cells were transplanted into L3-4, L4-5 and L5-6, respectively. RESULTS AND CONCLUSION: (1) Four weeks after the cell transplantation, the signal intensity of T2 weighted images in the L3-4 (non-transfection group) and L5-6 (blank plasmid group) discs was increased by magnetic resonance imaging, and the significant difference was found in L4-5 discs (shRNA-Notch-1 plasmid group) in comparison with other two groups (P < 0.05). (2) Toluidine blue staining showed that, the expression of proteoglycan in the L4-5 discs (shRNA-Notch-1 plasmid group) was significantly higher than that in L3-4 (non-transfection group) and L5-6 (blank plasmid group) discs. (3) Reverse transcription-polymerase chain reaction and western blot assay showed that, the expression of collagen II and proteoglycan mRNA and protein in the L4-5 discs (shRNA-Notch-1 plasmid group) were significantly increased compared with L3-4 (non-transfection group) and L5-6 (blank plasmid group) discs. Transplantation of bone marrow mesenchymal stem cells of Notch-1 gene knockout rabbits can effective restore the degenerated intervertebral discs. Figures and Tables | References | Related Articles | Metrics

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Dynamically observing chondrogenic differentiation of bone marrow mesenchymal stem cells in vitro Feng Jun-wei, Wang Yue, Lü Bo, Hao Peng, Tang Liu-yi, Zhu Jian-xin, Zhu Zong-dong, Tan Bo 2013, 17 (36):  6409-6416.  doi: 10.3969/j.issn.2095-4344.2013.36.005 Abstract ( 283 )   PDF (2588KB) ( 418 )   Save BACKGROUND: The reported time of bone marrow mesenchymal stem cells induced to differentiate into chondrocytes is different. Few studies have observed and compared the cells’ dynamic transformation during the induction process. OBJECTIVE: To observe the dynamic differentiation and the mature time of rabbit bone marrow mesenchymal stem cells which were directionally induced to chondroblasts for 8, 11, 14, 17, 20 days. METHODS: Bone marrow was aspirated from the femur of New Zeal rabbits, and bone marrow mesenchymal stem cells were isolated by gradient centrifugation. After cultivation and amplification, bone marrow mesenchymal stem cells at passage 3 were directionally induced to chondrocytes by the serum-free medium containing transforming growth factor beta-1. The experiments were divided into five groups according to different induction time points: 8 days, 11 days, 14 days, 17 days, 20 days. Then cellular morphology, toluidine blue staining, type Ⅱ collagen immunohistochemistry, aggrecan content in induction medium, and chondrogenic differentiation in each group were observed and compared. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells had apparently transformed in morphology at 8 days of induction, and presented obvious chondrocytes’ morphology at 14 days. The aggrecan in induction medium could be detected at a low level at 4 days, significantly increased at 8 days, and maintained slow increasing at 20 days. At 14 days, the metachromatic particles could be found by toluidine blue staining, and the collagen type Ⅱ immunohistochemistry was significantly positive in cell climbing slice. Experimental findings indicate that, bone marrow mesenchymal stem cells that are monolayer cultured in a high density can be induced into chondroblasts at the effect of transforming growth factor beta-1 and other factors. There are a few chondroblasts in the early induction process, then cells begin to have chondrocytes morphology and function after induced for 8 days, and may differentiate to mature chondrocytes at 14 days. In addition, they can keep a high biological activity in the induction process. Figures and Tables | References | Related Articles | Metrics

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Supernatant of myocardiocyte induces differentiation of bone marrow-derived mesenchymal stem cells Li Chun-mei, Wang Xiu-li, Zhu Hai-liang, Wang Jie, Gong Hai-bin 2013, 17 (36):  6417-6422.  doi: 10.3969/j.issn.2095-4344.2013.36.006 Abstract ( 244 )   PDF (1538KB) ( 419 )   Save BACKGROUND: Culture supernatant containing myocardiocyte has been demonstrated to induce differentiation of bone marrow-derived mesenchymal stem cells into myocardiocyte-like cells. This may associate with some or several cytokines in the culture supernatant. OBJECTIVE: To explore if the supernatant of myocardiocyte induces bone marrow-derived mesenchymal stem cells to differentiate into myocardiocyte-like cells is associated with the different cytokine content in the supernatant of myocardiocyte. METHODS: Bone marrow-derived mesenchymal stem cells were isolated and cultured in vitro by the whole bone marrow adherent culture. Cardiocytes were isolated and cultured by enzyme digestion. Bone marrow-derived mesenchymal stem cells (1×108/L) and cardiocytes (1×105/L) were cultured for 72 hours and the supernatant was collected. Hepatocyte growth factor, insulin-like growth factor 1, platelet-derived growth factor, stem cell factor, fibroblast growth factor and vascular endothelial growth factor levels in culture supernatant of bone marrow-derived mesenchymal stem cells and cardiocytes were detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: The content of insulin-like growth factor 1, platelet-derived growth factor and fibroblast growth factor in the supernatant of cardiocytes was significantly higher in cardiocytes group compared with bone marrow-derived mesenchymal stem cells group (P < 0.01). Results indicated that insulin-like growth factor 1, platelet-derived growth factor and fibroblast growth factor in the supernatant of cardiocytes may have capability to induce bone marrow-derived mesenchymal stem cells to differentiate into myocardiocyte-like cells, and insulin-like growth factor 1 may serve as the main cytokine. Figures and Tables | References | Related Articles | Metrics

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In vitro osteogenic differentiation of bone marrow mesenchymal stem cells from ovariectomied osteoporotic rats Wang Yun, Bao Xiao-ming, Hou Yong-xin, Li Jun, Zhang Min 2013, 17 (36):  6423-6429.  doi: 10.3969/j.issn.2095-4344.2013.36.007 Abstract ( 230 )   PDF (1915KB) ( 348 )   Save BACKGROUND: Cytological studies show that bone marrow mesenchymal stem cells play an important role in postmenopausal osteoporosis mechanism. OBJECTIVE: To study the osteogenic differentiation in vitro of bone marrow mesenchymal stem cells from ovariectomied osteoporotic rats.  METHODS: The osteoporotic animal model was established by performing ovariectomy in the 6-month-old female Sprague-Dawley rats. There were four groups: bone marrow mesenchymal stem cells control group, bone marrow mesenchymal stem cells osteoporosis group, bone marrow mesenchymal stem cells osteogenic induction group and oseogenesis induction group. Bone marrow mesenchymal stem cells were isolated from the rats of control group and oseogenesis induction group by means of the whole bone marrow adherence method and cultured to the 3rd generation. Then the bone marrow mesenchymal stem cells were used in all the experiments. Cell morphology was observed under the inverted phase contrast microscope, cell cycle and proliferation index of bone marrow mesenchymal stem cells were detected by flow cytometry. After osteogenic induction, the expression level of alkaline phosphatase was detected, and the fornation of calcium nodes of bone marrow mesenchymal stem cells were marked by alizarin red staining. RESULTS AND CONCLUSION: The cells in the osteogenic induction group and oseogenesis induction group had the morphology of osteobalsts, and the change of morphology of the cells in the oseogenesis induction group was relatively tardiness. The proliferation index in the control group was higher than that in the osteoporosis group (P < 0.05); expression level of alkaline phosphatase in the osteogenic induction group was significantly higher than that in the oseogenesis induction group (P < 0. 05), and the control group was significantly higher than the oseogenesis group (P < 0.05). The alizarin red staining of the cells in the osteogenic induction group was positive, while negative in the control group and the oseogenesis group; the staining in the osteogenic induction group was stronger than that in the oseogenesis induction group. These findings indicate that both the proliferative potential and the osteogenic potential of bone marrow mesenchymal stem cells from the ovariectomized osteoporotic rats are decreased, which may be related with the ostoeporosis pathogensis of ovariectomied rats. Figures and Tables | References | Related Articles | Metrics

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Effects of transportation conditions on survival rate of umbilical cord mesenchymal stem cells Wang Da-kun, Wang Zhi-hui, Guo Hong, Zhang Le-ling 2013, 17 (36):  6430-6435.  doi: 10.3969/j.issn.2095-4344.2013.36.008 Abstract ( 268 )   PDF (1685KB) ( 424 )   Save BACKGROUND: General assessment of viability of umbilical cord mesenchymal stem cells during transport is not considered at home and abroad.    OBJECTIVE: To explore the effects of different transportation conditions such as albumin and transport time on survival rate of umbilical cord mesenchymal stem cells.  METHODS: Umbilical cord mesenchymal stem cells cultured in vitro were divided into experimental and control groups. Each of group contained 3.15×109/L cells in 3 mL normal saline. The experimental group contained 1% albumin in dark environment. The control group included two subgroups: dark preservation group with the absence of albumin and light preservation group with the presence of albumin. Cell counting, trypan blue staining and cell survival rate were compared at 0, 2, 4, 8, and 24 hours.  RESULTS AND CONCLUSION: Compared with the control group, no cell loss was observed in experimental group (with the presence of albumin in the dark) and a 90% viability ratio was achieved at 8 hours. In the control group without albumin, loss rate reached 38.5% and survival rate reached 86% at 8 hours. Results revealed that 1% albumin predominantly improved cell survival rate in long-distance transport of umbilical cord mesenchymal stem cells. When transport time was more than 8 hours, cell loss rate increased and cell survival rate downregulated. Our experimental data demonstrated that umbilical cord mesenchymal stem cells preserved in normal saline consisting of 1% albumin placing in dark environment at 16 ℃ apply to clinical application criterion in 8 hours. Figures and Tables | References | Related Articles | Metrics

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Differentiation of cryopreserved umbilical cord mesenchymal stem cells into osteoblasts Chen Yan, Pan Li-jie, Yuan Jie, Li Tian-xia 2013, 17 (36):  6436-6442.  doi: 10.3969/j.issn.2095-4344.2013.36.009 Abstract ( 346 )   PDF (878KB) ( 573 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells are considered as novel seed cells in bone tissue engineering. Cryopreservation is an effective method for storing cells for a long time. OBJECTIVE: To explore whether umbilical cord mesenchymal stem cells of cryopreservation could be induced to differentiated into osteoblasts. METHODS: Mesenchymal stem cells were isolated from the Wharton’s jelly of human umbilical cord tissue by the tissue explant adherent method. Morphology of primitive cells was observed by inverted microscopy. Immunophenotypes and cell cycle of umbilical cord mesenchymal stem cells were measured using flow cytometry. After frozen storage for 6 months, the second passage of umbilical cord mesenchymal stem cells was thawed and subcultured to passage 12. Upon induction with osteogenic inductive medium, the osteogenic ability of passage 12 of umbilical cord mesenchymal stem cell was evaluated by alkaline phosphatase activity, the immunofluorescent analysis of osteocalcin and bone sialoprotein and the assay of alizarin red staining   separately. RESULTS AND CONCLUSION: Primary umbilical cord mesenchymal stem cells displayed a typical fibroblast-like morphology. Flow cytometry showed that the cultured cells expressed high levels of the mesenchymal stem cells surface markers CD73, CD105 and CD90, but did not express hematopoietic cells surface markers CD34 and CD45. The survival rate of umbilical cord mesenchymal stem cells after resuscitation was 90%. The cell cycle analysis indicated that 75% of the cells of passage 8 were in G0/G1 phase and 25% in S+G2M phase. Passage 12 cells treated with osteogenic inductive medium displayed a higher alkaline phosphatase activity compared with control cells (P < 0.01). Moreover, the cells, induced in osteogenic inductive medium, were positive for osteocalcin and bone sialoprotein staining and formed the mineralized nodules. Umbilical cord mesenchymal stem cells still maintain their biological characteristics after cryopreservation, and can be induced into osteoblasts with osteogenic inductive medium. Figures and Tables | References | Related Articles | Metrics

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Serum-free culture promotes rat adipose-derived stem cells differentiating into endothelial cells Guo Huan-shan, Yan Ling 2013, 17 (36):  6443-6448.  doi: 10.3969/j.issn.2095-4344.2013.36.010 Abstract ( 471 )   PDF (780KB) ( 445 )   Save BACKGROUND: There are few reports about the effect of serum-free culture on the differentiation of rat adipose-derived stem cells into vascular endothelial cells. OBJECTIVE: To investigate the isolation, serum-free culture of rat adipose-derived stem cells differentiating into vascular endothelial cells. METHODS: The rat adipose-derived stem cells were isolated from male Sprague-Dawley rats and expanded to the third passage by enzymatic digestion-adherent explants method. In the experimental group, rat adipose-derived stem cells were cultured in serum-free medium for 24 hours. In the control group, rat adipose-derived stem cells were cultured in low-glucose Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum. After that, the cells were cultured in inducing medium for 3 weeks to differentiate into vascular endothelial cells. RESULTS AND CONCLUSION: The rat adipose-derived stem cells grew as polygonal or fusiform-shaped adherent cells when cultured in vitro, which could stably proliferate and passage. The rat adipose-derived stem cells showed very low expression of CD31, a cell surface marker, after passages. After directional differentiations into vascular endothelial cells, the cells were pebble-shaped under the inverted microscope. Expression of CD31 was up-regulated, which was much higher in the experimental group than the control group. The induced cells in the experimental group had stronger abilities than those in the control group to swallow Dil-labeled acetylated low-density lipoprotein and form tube-like structures on the matrigel after differentiation into vascular endothelial cells. So, rat adipose-derived stem cells could be highly successfully induced to differentiate into vascular endothelial cells in vitro after serum-free culture. Figures and Tables | References | Related Articles | Metrics

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Peripheral blood mononuclear cell transplantation for liver cirrhosis Zhu Ying, Lang Shuai, Cong Qing-wei, Zhao Gang 2013, 17 (36):  6449-6454.  doi: 10.3969/j.issn.2095-4344.2013.36.011 Abstract ( 297 )   PDF (764KB) ( 338 )   Save BACKGROUND: Compared with bone marrow transplantation, peripheral blood stem cell transplantation has its own advantages, including rich resources of stem cells from the peripheral blood, convenient and easy collection, without anesthesia, small trauma, easily accepted, high safety, and easy to restore the patient’s hematopoietic system. OBJECTIVE: To observe the function and safety of autologous peripheral blood mononuclear cells in the treatment of patients with decompensated cirrhosis. METHODS: Four patients with decompensated liver cirrhosis were selected from November 2010 to July 2011 in the First Affiliated Hospital of Dalian Medical University, aged 31-67 (averagely 44 years). Among them, three cases had hepatitis B, and another one had autoimmune liver disease. Peripheral blood stem cells were collected after being mobilized by granulocyte colony stimulating factor. Then, autologous peripheral blood stem cells were transplanted via a hepatic artery catheter. RESULTS AND CONCLUSION: There were no adverse reactions such as fever, bleeding and nausea after peripheral blood stem cell collection and hepatic artery transplantation. Symptoms such as fatigue, poor appetite and abdominal distension gradually improved at 1, 3 and 6 months after transplantation. Liver function and liver fibrosis indexes were improved to some extent after transplantation. Figures and Tables | References | Related Articles | Metrics

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Hepatic arterial transplantation of autologous bone marrow mesenchymal stem cells in treatment of decompensated liver cirrhosis Ouyang Shi, Liu Shu-ren, Cheng Tao, Chen Yang-shu, Kong Xiang-ping, Zhou Chi-long, Mu Liang-jing 2013, 17 (36):  6455-6461.  doi: 10.3969/j.issn.2095-4344.2013.36.012 Abstract ( 282 )   PDF (777KB) ( 825 )   Save BACKGROUND: Autologous bone marrow mesenchymal stem cells can treat decompensated liver cirrhosis, however, little evidence has addressed the controlled clinical research in hepatitis B patients with decompensated live cirrhosis. OBJECTIVE: To evaluate the clinical efficacy and safety of autologous bone marrow mesenchymal stem cells in the treatment of hepatitis B with decompensated live cirrhosis. METHODS: A total of 67 hepatitis B patients with decompensated live cirrhosis were divided into two groups according to their wishes to receive stem cell transplantation. The control group (34 patients) only received oral administration of nucleoside analog antivirus and supportive treatment. The treatment group (33 patients) received autologous bone marrow mesenchymal stem cells transplantation via hepatic artery plus antivirus and supportive treatment. The liver functional index, clinical signs and symptoms, adverse reactions were observed and compared at 4, 12, 24 weeks after treatment. RESULTS AND CONCLUSION: After treatment, all patients’ symptoms were improved to varying degrees. After 4 weeks of treatment, the liver functional indexes were all significantly improved compared with before treatment, the levels of alanme aminotransferase, cholinesterase and prothrombin activity in treatment group were significantly ameliorated compared with control group (P < 0.05). At 12 and 24 weeks of treatment, the alanme aminotransferase, albumin, total bilirubin, cholinesterase and prothrombin activity in control group and treatment group showed statistically significant differences compared with before treatment (P < 0.05). At the same time point, all the indicators in the treatment group were significantly ameliorated compared with control group (P < 0.05). The Child-pugh score and model for end-stage liver disease score declined at 4, 12, 24 weeks after treatment, showing significant differences compared with before treatment. The difference was also significant at the same time point between two groups. The treatment of nucleoside analogue antivirus combined with autologous bone marrow mesenchymal stem cells transplantation on hepatitis B patients with decompensated liver cirrhosis is an effective method to improve liver function and blood coagulation function, with symptom improvement, safety and low risk. Figures and Tables | References | Related Articles | Metrics

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Immunosuppression of bone marrow mesenchymal stem cells on T cells in patients with aplastic anemia Zhang Le-qin, Xiao Yang, Jiang Zu-jun, Xiao Hao-wen, Li Li, Pang Yan 2013, 17 (36):  6462-6467.  doi: 10.3969/j.issn.2095-4344.2013.36.013 Abstract ( 295 )   PDF (842KB) ( 409 )   Save BACKGROUND: To date, there are few domestic reports about the influence of bone marrow mesenchymal stem cells on T cells proliferation in patients with aplastic anemia. And no study addresses the topic that if bone marrow mesenchymal stem cells achieve immune regulation in aplastic anemia patients through inhibiting T cells proliferation. OBJECTIVE: To explore the effects of human bone marrow mesenchymal stem cells on T cells immune regulation in patients with aplastic anemia. METHODS: Human bone marrow mesenchymal stem cells were isolated, cultured and subcultivated in vitro. The morphological appearance of bone marrow mesenchymal stem cells was observed and surface markers were measured by flow cyometry. The bone marrow mesenchymal stem cells were co-cultured with T cells extracted from peripheral blood of healthy volunteers and aplastic anemia patients for 7 days. The expressions of interferon-γ, interleukin-4 and interleukin-10 in the supernatants were detected with enzyme linked immunosorbent assay. RESULTS AND CONCLUSION: The levels of interleukin-2 and interferon-γ in the supernatant of aplastic anemia patients were significantly higher (P < 0.05), while levels of interleukin-4 and interleukin-10 were significantly lower than that in healthy controls (P < 0.05). Bone marrow mesenchymal stem cells suppressed the elevated levels of interleukin-2 and interferon-γ, and enhanced the decreased levels of interleukin-4 and interleukin-10, thus regulating the immune dysfunction of aplastic anemia patients. Figures and Tables | Related Articles | Metrics

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Bone marrow mesenchymal stem cells against monocrotaline-induced pulmonary artery hypertension Lu Yan, Kong Ling-cai, Zhang Zhao-hua 2013, 17 (36):  6468-6473.  doi: 10.3969/j.issn.2095-4344.2013.36.014 Abstract ( 362 )   PDF (490KB) ( 412 )   Save BACKGROUND: Stem cell transplantation has a certain effect in the treatment pulmonary arterial hypertension. OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells transplantation on the treatment of pulmonary arterial hypertension and to discuss the mechanism. METHODS: Bone marrow mesenchymal stem cells were in vitro cultured, purified and amplified by density gradient centrifugation method, and labeled with the fluorescent dye for preparation. Pulmonary arterial hypertension model was established by subcutaneous injection of monocrotaline. One week after modeling, the rats were randomly divided into three groups. Rats in the stem cell transplantation group and pulmonary arterial hypertension group received subcutaneous injection of monocrotaline to establish the pulmonary arterial hypertension model. One week later, the rats in the stem cell transplantation group received sublingual vein injection of bone marrow mesenchymal stem cell solution, the rats in the pulmonary arterial hypertension group were injected with the culture medium without stem cells, and the rats in the control group were injected with the normal saline in the same dose. RESULTS AND CONCLUSION: At 2 weeks after transplantation, compared with the mesenchymal-induced pulmonary arterial hypertension rats, the hemodynamic parameters and the ratio of right ventricular/body weight of the rats in the stem cell transplantation group were significantly improved (P < 0.05); the degree of pulmonary vascular remodeling was significantly reduced (P < 0.05). Fluorescence microscope observation showed that the transplanted bone marrow mesenchymal stem cells could alive at least 2 weeks in the stem cell transplantation group, and part of the stem cells could differentiate into pulmonary vascular endothelial cells. The results show that bone marrow mesenchymal stem cell transplantation can significantly improve the pulmonary vascular and right ventricular structural impairments in the rats with mesenchymal-induced pulmonary arterial hypertension. Figures and Tables | References | Related Articles | Metrics

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Syngeneic and allogeneic fetal liver stem cell transplantation in the treatment of mouse hepatic cirrhosis Han Bo, Xu San-rong, Zhang Jin, Zhou Qing, Li Wei 2013, 17 (36):  6474-6480.  doi: 10.3969/j.issn.2095-4344.2013.36.015 Abstract ( 430 )   PDF (2021KB) ( 657 )   Save BACKGROUND: Immunity of fetal liver stem cell transplantation is rarely reported, syngeneic and allogeneic fetal liver stem cell transplantation in the treatment of hepatic cirrhosis is still unclear. OBJECTIVE: To observe the therapeutic effects of syngeneic and allogeneic fetal liver stem cell transplantation on hepatic cirrhosis as well as immune rejections during the therapeutic process. METHODS: The fetal liver stem/progenitor cells from BALB/c and C57BL/6 mice were isolated and purified by the type IV collagen enzyme digestion method. A total of 104 healthy BALB/c mice were randomly assigned to four groups. Normal control group: no treatment; Hepatic cirrhosis group, syngeneic transplantation group and allogeneic transplantation group: 16 weeks after hepatic cirrhosis models of mice were developed by intraperitoneal injection with carbon tetrachloride, physiological saline, syngeneic fetal liver stem cells and allogeneic fetal liver stem cells were injected via the caudal vein. Finally, the survival statuses, liver function, hepatic fibrosis index, the number and ratio of immune cells (CD4+T, CD8+T, NK, NKT) and histopathologic examinations were compared in each group after transplantation 4 weeks. RESULTS AND CONCLUSION: The survival rates in the two transplantation groups were both 100%, which was significantly higher than that in the hepatic cirrhosis group (67%, P < 0.05). The liver function and liver fibrosis index in each group did not show statistical differences (P > 0.05). Immunological tests showed no difference between groups (P > 0.05). Pathohistology examination of hepatic tissue repair: Allogeneic transplantation group > syngeneic transplantation group > hepatic cirrhosis group. Hence, fetal liver stem cell transplantation via the caudal vein could elevate the survival rate of hepatic cirrhosis mice, alleviate the degree of hepatocyte necrosis. There is no immunologic rejection during syngeneic and allogeneic fetal liver stem cell transplantation that could help to treat hepatic cirrhosis in mice. Figures and Tables | References | Related Articles | Metrics

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Molecular imaging for tracking transplanted embryonic stem cells in the treatment of acute liver injury Yao Xin-peng, Xu Yang, Zhang Lu, Leng Liang, Su Wei-jun, Wang Li-na, Tong Ling-ling, Li Zong-jin, Kong De-ling 2013, 17 (36):  6481-6488.  doi: 10.3969/j.issn.2095-4344.2013.36.016 Abstract ( 347 )   PDF (3819KB) ( 447 )   Save BACKGROUND: Embryonic stem cells have the capacity of multi-differentiation potential, and have been utilized for the therapy of acute liver injury. However, the migration and proliferation of embryonic stem cells after transplantation remains not well characterized. OBJECTIVE: To track the transplanted embryonic stem cells in repairing acute liver injury by bioluminescence imaging technology. METHODS: Murine embryonic stem cells (D3) were transducted with a construct composed of firefly luciferase, monomeric red fluorescence protein and herpes simplex virus truncated thymidine kinase triple fusion reporter genes by lentivirus system. Stable D3 embryonic stem cells integrating three report genes were screened. The undifferentiated embryonic stem cells or differentiated embryonic stem cells from the 6-day-old embryoid body were transplanted into acute liver injury model of SV129 mouse through spleen, and the transplanted cells were monitored by bioluminescence imaging technology. RESULTS AND CONCLUSION: Reverse transcription PCR results showed that the expression level of Oct-4 and Nanog was not affected in embryonic stem cells transducted with triple fusion reporter gene compared with wild-type embryonic stem cells. The migration process of transplanted cells was visualized by bioluminescence imaging technology. Teratomas were found in both triple fusion-embryonic stem cells treatment group and triple fusion-embryoid body cells treatment group at liver, and the teratoma formation could be suppressed by ganciclovir administration because ganciclovir can react with herpes simplex virus truncated thymidine kinase and trigger cell necrosis process. Histological analysis showed that teratomas comprised tissues from all three germ layers. These results demonstrate that triple gene fusion does not affect differentiation potential of embryonic stem cells and it is risky to utilize embryonic stem cells for cell therapy, because it affects repair of liver injury. The therapy strategy requires further improvement and real-time visualizing of embryonic stem cells in vivo is absolutely necessary. Figures and Tables | References | Related Articles | Metrics

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Quantitative analysis of SOX9 and type Ⅱ collagen mRNA in the three-lineage differentiation of rat mesenchymal stem cells Liang Da-chuan, Bai Jie-yu, Du Shao-hua, Cheng Peng, Kang Ning, Wang Zhen, Huang Qiang-kai, Yang Zi-quan 2013, 17 (36):  6489-6494.  doi: 10.3969/j.issn.2095-4344.2013.36.017 Abstract ( 300 )   PDF (1828KB) ( 425 )   Save BACKGROUND: The main component of cartilage, type Ⅱ collagen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner. OBJECTIVE: To observe the variation of SOX9 and type Ⅱ collagen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cells, and to explore the correlation of SOX9 expression and type Ⅱ collagen. METHODS: Bone marrow mesenchymal stem cells were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cell phenotype was identified with flow cytometry. Cells were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cells were used as a control group. The total RNA of cells was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ collagen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cells were stained by immunofluorescence to observe the differentiation and perform statistical analysis.      RESULTS AND CONCLUSION: Passage 3 bone marrow mesenchymal stem cells grew well, and cell phenotype was confirmed as stem cells by flow cytometry. The staining results showed that, the cells differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cells were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ collagen mRNA levels in the induced cells were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at  3 and 7 days, and then decreased at 14 days. While type Ⅱ collagen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ collagen expression gradually decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P > 0.05), while type Ⅱ collagen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱ collagen, which may alter along with the SOX9 in the early chondrogenic differentiation; SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation. Figures and Tables | Related Articles | Metrics

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Phenotypes and characteristics of human skin-derived precursors Liu Gui-ying, Yang Li-ye, Li Wen-yu, Zheng Jia-kun 2013, 17 (36):  6495-6500.  doi: 10.3969/j.issn.2095-4344.2013.36.018 Abstract ( 468 )   PDF (2338KB) ( 372 )   Save BACKGROUND: Human skin-derived precursors can be cultured for a long term in vitro, and differentiated into neurons, glial cells, smooth muscle cells, Schwann cells and cells with peripheral neurons phenotype. OBJECTIVE: To investigate the culture conditions and multiple differentiation capacity of multipotential stem cells from human skin, especially the potentials of differentiating into neurons and osteoblasts. METHODS: Human skin-derived precursor cells were cultured with trypsin digestion method, and identified with immunocytochemistry. Cells at passages 3-4 were induced to differentiate into neurons and osteoblasts, and underwent von Kossa staining protocol for calcium, chondrocyte induction, toluidine blue staining, immunohistochemical staining and Sudan black staining. The expression of nestin, vimentin, βIII-tubulin, S100 and collagen II in the human skin-derived precursors was detected. RESULTS AND CONCLUSION: The human skin-derived precursor cells cultured with trypsin digestion method could proliferate and form suspending spheres, and nestin positive cells were detected at any time point of the culture. All the cultured cells expressed vimentin, and some adherent cells expressed βIII-tubulin. Human skin-derived precursor cells were induced with Salvia miltiorrhiza to differentiate into neuron-like cells, and expressed marker of nerve cells. Skin-derived precursors could be induced to differentiate into osteoblasts and von Kossa staining displayed black calcified nodules in the culture dish. Skin-derived precursors could also be induced to differentiate into chondrocytes, and toluidine blue staining was strongly positive, and some cells expressed collagen II, which suggested that, the differentiated cells contained chondrocytes. Experimental findings indicate that, skin contains multipotential stem cells that are capable of differentiating into osteoblasts, chondrocytes, Schwann cells and oligodendrocytes. Figures and Tables | References | Related Articles | Metrics

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Platelet-rich plasma affects the proliferation and collagen production of mesenchymal stem cells Rong Chun, Shen Yan-qing, Lü Ya-qing, Li Ming-yu, Xia Chang-suo 2013, 17 (36):  6501-6507.  doi: 10.3969/j.issn.2095-4344.2013.36.019 Abstract ( 275 )   PDF (357KB) ( 807 )   Save BACKGROUND: Mesenchymal stem cells are the seed cells for tendon tissue engineering which can be obtained in large quantities, but how to induce in vitro is a key technology. OBJECTIVE: To explore the effect of platelet-rich plasma on the proliferation and collagen production of in vitro cultured mesenchymal stem cells. METHODS: The rabbit mesenchymal stem cells were separated ad cultured. The high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group were set to induce the mesenchymal stem cells, and the blank control group was set as control.    RESULTS AND CONCLUSION: The proliferation of mesenchymal stem cells in the high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group was high, and in rapid growth with big increase amplitude, and there was no significant difference in the proliferation when compared with the blank control group (P < 0.05). The effect was positively correlated with the culture time, and after cultured for a certain time, the effect was in dose-dependent manner, as higher dose platelet-rich plasma had more significant effect on the proliferation of the cells. The results indicate that platelet-rich plasma can significantly promote the synthesis of collagen type Ⅰ and Ⅲ of mesenchymal stem cells, the higher the dose, the more significant the effect on the collagen. Figures and Tables | References | Related Articles | Metrics

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Isolation, culture and characterization of endothelial progenitor cells from the human peripheral blood Qiao Wei, Ran Feng, Liu Chang-jian 2013, 17 (36):  6508-6514.  doi: 10.3969/j.issn.2095-4344.2013.36.020 Abstract ( 764 )   PDF (2271KB) ( 838 )   Save BACKGROUND: Endothelial progenitor cells, known as the precursor cells of mature endothelial cells, have the function of neovascularization and neoendothelialization. Therefore, endothelial progenitor cells have potential applicability in many fields. Endothelial progenitor cells can be isolated and cultured from different resources with different methods, but the biological properties and identification of endothelial progenitor cells still have controversies. OBJECTIVE: To explore the methods of isolation and culture of endothelial progenitor cells from the human peripheral blood and to identify the biological features of endothelial progenitor cells.  METHODS: Mononuclear cells were isolated from the human peripheral blood using density gradient centrifugation, and the cells were resuspended in endothelial basal medium-2 supplemented with the EGM-2-MV-SingleQuots. Then, the cells were inoculated in human fibronectin-coated culture flasks and cultured in EBM-2MV medium. The morphology of endothelial progenitor cells was observed. The proliferation potential and surface markers of endothelial progenitor cells were characterized carefully. Furthermore, the functional properties such as nitric oxide release and tube formation on Matrigel were also evaluated. RESULTS AND CONCLUSION: While adherent cells maintained, spindle-shaped cells formed a cell cluster after 6-7 days. Then, adherent cells developed to endothelial progenitor cells with a cobblestone appearance after 2-3 weeks. The endothelial progenitor cells were confluent with an outgrowth appearance. Endothelial progenitor cells had a higher proliferation potential compared with human aortic endothelial cells under the same culture condition. Endothelial progenitor cells expressed CD31, CD34, CD144 and KDR, displaying an obvious endothelial phenotype. Endothelial progenitor cells were also found to uptake DiL-acLDL and exhibit lectin binding capability. Furthermore, endothelial progenitor cells were able to form capillary tubes on Matrigel and had the ability to release nitric oxide. Therefore, endothelial progenitor cells can be obtained from the human peripheral blood by density gradient centrifugation and adherent culture. A combining method for the identification of endothelial progenitor cells should be recommended. Figures and Tables | References | Related Articles | Metrics

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Combination of tanshinone IIa and astragaloside induces bone marrow mesenchymal stem cells differentiating into myocardium-like cells Song Xue-yun, Long Yun, Tan Chao, Zheng Jing-hui 2013, 17 (36):  6515-6520.  doi: 10.3969/j.issn.2095-4344.2013.36.021 Abstract ( 311 )   PDF (2291KB) ( 421 )   Save BACKGROUND: Astragaloside and tanshinone IIa are the effective components of traditional Chinese medicine treatment of myocardial ischemia, and its role in bone marrow mesenchymal stem cells differentiation into myocardium-like cells remains unclear. OBJECTIVE: To investigate the effect of tanshinone IIa and astragaloside on the differentiation of bone marrow mesenchymal stem cells into myocardium-like cells. METHODS: The maximal non-toxic concentrations of tanshinone IIa and astragaloside were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, to define the dose of the two in the induced differentiation of bone marrow mesenchymal stem cells into myocardium-like cells. The isolated and purified bone marrow mesenchymal stem cells were divided into five groups: astragaloside group, tanshinone IIa group, astragaloside + tanshinone IIa group, 5-azacitidine group, and blank control group. The expression of gap junction connexin 43 and troponin was determined with enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: The expression of gap junction connexin 43 and troponin in astragaloside group, tanshinone IIa group, astragaloside + tanshinone IIa group, 5-azacitidine group was higher than that in blank control group (P < 0.01). The astragaloside + tanshinone IIa group showed a higher expression of gap junction connexin 43 and troponin than astragaloside group and tanshinone IIa group (P < 0.01). There was no significant difference in the expression of gap junction connexin 43 and troponin between astragaloside + tanshinone IIa group and 5-azacitidine group (P > 0.05). A combined use of astragaloside and tanshinone IIa can induce bone marrow mesenchymal stem cells to differentiate into myocardium-like cells, and their joint role is better than the role of a single ingredient. Figures and Tables | References | Related Articles | Metrics

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Impact of mesenchymal stem cells on the proliferation, invasion and biological behaviors of hepatocellular carcinoma cells Zheng Peng-xiang, Zhou Huan, Tan Jian-ming 2013, 17 (36):  6521-6526.  doi: 10.3969/j.issn.2095-4344.2013.36.022 Abstract ( 285 )   PDF (2338KB) ( 480 )   Save BACKGROUND: Liver cancer pathogenesis and intervention have attracted increasing attentions. Mesenchymal stem cells become a popular tool for cell cancer research, because of their low immunogenicity and tumor tropism. At present, mesenchymal stem cells have been applied to the study of liver cancer. OBJECTIVE: To summarize the advances of mesenchymal stem cells used in liver cancer in basic and clinical research. METHODS: An online retrieval of CNKI and Pubmed database was performed by the first author for the articles about mesenchymal stem cells and effect of modified mesenchymal stem cells on hepatoma carcinoma cells published from January 2004 to January 2013. The key words were “mesenchymal stem cell, liver cancer, tumor” in Chinese and English. Repetitive research was excluded, and 47 studies met the inclusion criteria. RESULTS AND CONCLUSION: Stem cells are seldom reported in liver cancer, and the limited present study show that mesenchymal stem cells may have a certain influence on the hepatoma carcinoma cell proliferation, invasion and biological behavior. However, due to the differences of cell lines used by the various laboratories, experimental conditions, animal models, as well as infusion means of stem cells, experimental results are also inconsistent. Scholars have conducted a series of studies on the mechanism of the Wnt pathway and tumor necrosis factor-related apoptosis-inducing ligand pathway. Tropism of mesenchymal stem cells to tumor cells, including liver cancer is widely recognized, so scholars imported therapeutic genes and drugs into mesenchymal stem cells to interfere with the development of liver cancer, and have achieved some progress. This evidence provides new avenues for cell therapy for liver cancer. Less safety studies in vivo and clinical trials of mesenchymal stem cells are available, therefore security risks deserve further research. Figures and Tables | References | Related Articles | Metrics

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Biological markers of epidermal stem cells Hao Fei, Zhang Ling, Wu Yan 2013, 17 (36):  6527-6532.  doi: 10.3969/j.issn.2095-4344.2013.36.023 Abstract ( 515 )   PDF (2555KB) ( 957 )   Save BACKGROUND: Repair of large-area burns and severe post-traumatic skin defects has always been urgent clinical breakthrough technology bottleneck. With the development of tissue engineering, epidermal stem cells are increasingly being used in tissue engineering, cell replacement therapy and genetic engineering. Therefore, the isolation and identification of epidermal stem cells is becoming the research focus of concern. OBJECTIVE: To review the research progress in biological markers of epidermal stem cells. METHODS: The Chinese Biomedical Literature database, CNKI database, China Academic Journals Full-text database, PubMed database and EMbase database were retrieved for articles about specific markers of epidermal stem cells using the keywords of “epidermal stem cells, integrin, keratin, P63, CD71, telomerase, ACE, cx43, hoechst” in Chinese and English. Older theoretical perspectives and repetitive research were excluded. RESULTS AND CONCLUSION: Finally, only 40 articles were included in result analysis. Epidermal stem cells bring a new source for skin tissue repair. Epidermal stem cells distribute in the follicle eminence and basal layer of the epidermis. About 4% cells in the basal layer, however, are stem cells. Therefore, it is critical to correctly isolate, culture and identify skin stem cells. As a reason, specific markers of epidermal stem cells become a hotspot. Currently, a great progress in the biological markers of epidermal stem cells has been made, but there is still no absolute and proven marker for epidermal stem cells. Most studies are focusing on integrin, keratin, P63, CD71, connexin 43, and telomerase. In addition, hoechst, CD90, CD98, CD200 have been reported recently. Each marker has its own shortcomings, and there are still many problems that need to be solved. Figures and Tables | References | Related Articles | Metrics

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Insight into induced pluripotent stem cells in retinal diseases Deng Jun, Chen Xiao, Yan Ying, Zhou He-zheng 2013, 17 (36):  6533-6540.  doi: 10.3969/j.issn.2095-4344.2013.36.024 Abstract ( 486 )   PDF (3249KB) ( 863 )   Save BACKGROUND: Retinal diseases are caused by damage or apoptosis of the retinal cells and lack of ideal treatment. Induced pluripotent stem cells have self-renewal ability and differentiation potential. A new potential source from which to produce stem cells as a therapeutic platform for the treatment of retinal diseases and avoid ethics issues is the research focus currently. OBJECTIVE: To review recent advancement of induced pluripotent stem cells and the potential utility for retinal diseases. METHODS: The first author searched related articles published from 2006 to 2011 by computer retrieval in PubMed database using the keywords of “induced pluripotent stem cells, retinal, photoreceptor, retinal pigment epithelium, retinal ganglion cell” in English. Finally, totally 42 papers were involved. RESULTS AND CONCLUSION: Induced pluripotent stem cells can be induced to differentiate into retinal cells which have a possibility to be used as donor cells for transplantation therapies. Patient-derived induced pluripotent stem cells can be used as disease models for mechanistic studies and drug screening. It is a promising therapy for retinal degenerative diseases. However, some problems still remain, such as risk of teratoma formation and low efficiency of somatic cells to be successfully induced into induced pluripotent stem cells. It still requires further more studies. Figures and Tables | References | Related Articles | Metrics