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18 February 2017, Volume 21 Issue 5 Previous Issue    Next Issue

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Quercetin inhibits proliferation and promotes apoptosis of glioma stem cells via the STAT pathway Liu Yi-feng, Zhang Bao-chao, Wen Chang-ming, Wen Gong-ling, Zhou Gou-ping, Zhang Jing-wei, He Hai-fa, Wang Ning, Li Wei 2017, 21 (5):  657-662.  doi: 10.3969/j.issn.2095-4344.2017.05.001 Abstract ( 287 )   PDF (1049KB) ( 222 )   Save BACKGROUND: Several studies have reported that quercetin can inhibit the proliferation and migration but promotes apoptosis of tumor cells. OBJECTIVE: To explore the effect of quercetin on the proliferation and apoptosis of glioma stem cells and the signal pathway involved. METHODS: Glioma stem cells were isolated by immunomagnetic beads and treated in culture medium containing different concentrations of quercetin (0, 25, 50, 100 μmol/L). Cell proliferation was measured by MTT and apoptosis measured by flow cytometry at 48 hours after culture. The expression levels of Bcl-2, Bax and survivin, which are related to apoptosis, were detected by western blot. The expression of proliferating cell nuclear antigen (PCNA), which is related to proliferation, was also detected by western blot. The expression of STAT3 and p-STAT3 was also determined by western blot. RESULTS AND CONCLUSION: (1) Compared with the control (0 μmol/L) group, quercetin inhibited the proliferation of glioma stem cells in a dose-depended manner (P < 0.05). With the increase of the concentration of quercetin, the expression of PCNA was increased (P < 0.05). (2) Quercetin induced apoptosis of glioma stem cells dose-dependently (P < 0.05). With the increase of the concentration of quercetin, the expression of Bcl-2 and survivin was decreased, while the expression Bax was increased. (3) Quercetin could also inhibit phosphorylation of STAT3 dose-dependently, but the level of STAT3 was not changed. To conclude, these results show that quercetin could inhibit the proliferation of glioma stem cells and promote apoptosis via the STAT pathway.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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TRAP1 gene silencing effect on biological properties of CD24-CD44+ human laryngeal squamous cell carcinoma stem cells Su Jing, Xue Hai-tao, Tian Jun-hai, Zhang Ji-hua 2017, 21 (5):  663-668.  doi: 10.3969/j.issn.2095-4344.2017.05.002 Abstract ( 365 )   PDF (1319KB) ( 192 )   Save BACKGROUND: Studies have indicated that the abnormal expression of tumor necrosis factor receptor-associated protein 1 (TRAP1) is closely related to the occurrence and development of a variety of tumors. Therefore, targeted inhibition of TRAP1 expression has become an important target for the treatment or intervention of tumor growth. OBJECTIVE: To explore the effect of the TRAP1 gene silencing on the proliferation and apoptosis of human laryngeal squamous cell carcinoma stem cells. METHODS: CD24-CD44- human laryngeal squamous cell carcinoma stem cells were isolated by flow cytometry. Interfering RNA (siRNA) sequences for small molecule TRAP1 gene was designed and transferred into human laryngeal cancer stem cells by LipofectamineTM 2000. Flow cytometry, MTT assay, cell clone formation assay and TUNEL apoptosis assay were used to evaluate the effect of silencing TRAP1 gene on the proliferation and apoptosis of CD24-CD44+ laryngeal cancer stem cells. RESULTS AND CONCLUSION: Compared with CD24+CD44- cells, CD24-CD44+ cells upregulated OCT4, SOX2, NANOG and TRAP1 expression levels (P < 0.05). However, the expression of TRAP1 protein in human laryngeal squamous cell carcinoma was significantly decreased after RNA interference (P < 0.05). The growth rate of TRAP1 gene silenced human laryngeal squamous cell carcinoma was significantly reduced (P < 0.05), the cell arrest was in the G0/G1 phase, the number of cells in the S phase was decreased (P < 0.05), and there was no significant change in the M phase. TRAP1 gene silencing significantly inhibited the proliferation of human laryngeal squamous cell carcinoma stem cells (P < 0.05). Compared to the non-transfected cells, the TRAP1 gene silencing significantly reduced the clone formation ability of transfected human laryngeal squamous cell carcinoma stem cells (P < 0.05), and TRAP1 gene silenced-human laryngeal squamous cell carcinoma stem cells were more easy to trigger apoptosis by upregulating BAD and BAX expression levels (P < 0.05). Overall, our experimental results indicate that the specific interference of TRAP1 gene expression could inhibit the proliferation and promote apoptosis of human laryngeal squamous cell carcinoma stem cells.    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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A preliminary proof of kidney-deficiency phenotype in human bone marrow mesenchymal stem cells with estrogen deficiency Wei Qiu-shi, He Wei, Chen Zhen-qiu, Chen Da, Hong Guo-ju, Chen Jian-fa, Yang Peng 2017, 21 (5):  669-675.  doi: 10.3969/j.issn.2095-4344.2017.05.003 Abstract ( 235 )   PDF (6157KB) ( 183 )   Save BACKGROUND: As the presence of decreased proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs) results from estrogen deficiency-induced postmenopausal osteoporosis, it is of significance to study the relevant pathogenesis and treatment strategies from the perspective of estrogen and its receptors. OBJECTIVE: To observe the characteristics of kidney-deficiency phenotype in estrogen deficiency-induced BMSCs. METHODS: Passage 3 BMSCs were cultured in the osteogenic induction medium. The cells were divided into five groups, and cultured with different concentrations (0, 10-10, 10-9, 10-8, 10-7 mol/L) of letrozole. BMSCs with no osteogenic induction were used as blank controls. The expression levels of AROM were detected at mRNA and protein levels by real-time PCR and western blot methods. Cell proliferation of BMSCs was detected using MTT assay. Alkaline phosphatase activity assay was used to detect BMSCs osteogenic ability. ELISA was used to detect estradiol levels in cell supernatants. The expression of cAMP mRNA was detected by real-time PCR method. RESULTS AND CONCLUSION: 10-8 mol/L Letrozole exhibited the best effect to inhibit the expression of AROM mRNA and protein. After 72 hours of culture, the proliferation of BMSCs fell to the bottom in the 10-8 mol/L letrozole group. After 14 days of culture, the alkaline phosphatase activity in the 10-8 mol/L letrozole group was lower than that in the other groups. ELISA showed that estradiol level in supernatants was lower in the 10-8 mol/L letrozole group than the other groups. Real-time PCR showed that cAMP mRNA expression was lower in the 10-8 mol/L letrozole group than the other groups. These results suggest that 10-8 mol/L letrozole can significantly inhibit the proliferation and osteogenic differentiation of BMSCs, and can significantly decrease the expression of cAMP and the potential of synthesizing and secreting estradiol. Therefore, the process of AROM-mediated estradiol synthesis in BMSCs can be inhibited by 10-8 mol/L letrozole, and has the characteristics of kidney-deficiency phenotype.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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In vivo distribution of luciferase gene-labeled bone marrow mesenchymal stem cells infused through different approaches Sun Xiao-wei, Huang Hao, Zhou Yong-jun, Chen Xiao-li, Qiao Peng-xin, Zou Chun, Zhang Qiu-xia, Jiang Qian-li 2017, 21 (5):  676-681.  doi: 10.3969/j.issn.2095-4344.2017.05.004 Abstract ( 307 )   PDF (5556KB) ( 178 )   Save BACKGROUND: Most bone marrow mesenchymal stem cells are infused intravenously and have very low efficiency of homing to the bone marrow. However, cell infusion via the femoral approach is little reported. OBJECTIVE: To explore the distribution of luciferase gene modified red fluorescent protein transgenic bone marrow mesenchymal stem cells in vivo through different infusion routes. METHODS: Luciferase gene modified bone marrow mesenchymal stem cells at different gradients (5×106, 1×106, 1×105, 1×104) were seeded or injected into the in vitro pore plate or free femurs to observe the fluorescence imaging and select the best concentration of cells. Luciferase gene modified bone marrow mesenchymal stem cells at the best cell concentration were injected into the mice via the femur and the tail vein, respectively. The distribution of fluorescence and cell number in the mice were explored by using bioluminescence, pathological examination, flow cytometry and quantitative PCR. RESULTS AND CONCLUSION: Ex vivo fluorescence intensity of luciferase gene modified bone marrow mesenchymal stem cells was positively correlated with the cell concentration; fluorescent cells in vivo appeared in the femur first and then quickly spread to the lungs in the femur group, while fluorescent cells in the tail vein group spread to the lungs quickly after cell infusion. Fluorescent cells could be seen in the spleen, liver and other organs 24 hours later in the two groups. The distribution and migration of cells in mice could be observed successfully by bioluminescence; 5 minutes after cell infusion, the lungs of mice in the two groups began to emit fluorescence that could spread to the liver, spleen and other tissues 24 hours later, and the fluorescence intensity reached its peak after 15 minutes. The distribution of bone marrow mesenchymal stem cells in mice had no significant difference between the femur group and the tail vein group. To conclude, cell injection through the bone marrow cavity and tail vein fails to promote the homing of bone marrow mesenchymal stem cells to the bone marrow.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Isolation and culture of bone marrow mesenchymal stem cells and their effects on immune regulation of CD8+T lymphocytes Zhao Xiao-qiang, Shao Ming, Yang Hai-ping, Sun Ling 2017, 21 (5):  682-686.  doi: 10.3969/j.issn.2095-4344.2017.05.005 Abstract ( 276 )   PDF (3707KB) ( 281 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have immunomodulatory properties and have potential applications in immunosuppression. OBJECTIVE: To investigate the isolation and culture of bone marrow mesenchymal stem cells and the immunoregulation of CD8+T lymphocytes. METHODS: Primary bone marrow mesenchymal stem cells were isolated from Wistar rats by bone marrow adherence method. The primary cells were purified by differential adherence and digestion, and collagen type I was added as extracellular matrix to expand bone marrow mesenchymal stem cells. Passage 2 bone marrow mesenchymal stem cells at densities of 0, 1×103, 1×104 and 1×105/well were co-cultured with CD8+T lymphocytes, followed by phytohemagglutinin stimulation for 68 hours. T lymphocyte aggregation and proliferation were detected by staining with staining with 5,6-carboxyfluorescein diacetate succinimidyl ester and MTT, respectively. RESULTS AND CONCLUSION: Different concentrations of bone marrow mesenchymal stem cells had an inhibitory effect on T lymphocytes, but this effect was weakest for the cells at the density of 1×103 per well and strongest for the cells at the density of 1×105 per well. Obvious agglomeration was observed in the same media. The proliferation rates of CD8+T lymphocytes were (44.83±4.92)%, (31.94±6.28)% and (15.77±3.98)%, respectively, after co-culture with 1×103, 1×104, 1×105/well bone marrow mesenchymal stem cells. There were significant differences between groups (P < 0.05). These results showed that primary rat bone marrow mesenchymal stem cells could be obtained by direct adherence method of the whole bone marrow, and the cells could be purified by differential adherence combined with digestion and control method. Bone marrow mesenchymal stem cells could inhibit T lymphocyte proliferation in a concentration- dependent manner.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of bone marrow mesenchymal stem cells on MHCC97-H cells after transforming growth factor beta1 and osteopontin gene interference Li Tian-ran, Huang Xiao-bin, Huang Chu-heng, Lu Guang-ming, Li Yan-jun 2017, 21 (5):  687-692.  doi: 10.3969/j.issn.2095-4344.2017.05.006 Abstract ( 265 )   PDF (4309KB) ( 212 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are the focus of research on the proliferation and metastasis of hepatocellular carcinoma cells. By genetic engineering techniques, the hepatocellular carcinoma cells can be induced to reduce the expression of bioactive factors, thereby seeking suitable intervention targets for improving the interventional effect of BMSCs. OBJECTIVE: To silence the expression of transforming growth factor beta1 (TGFβ1) and osteopontin (OPN) in high metastatic potential hepatocellular carcinoma cells (MHCC97-H) followed by co-culture with BMSCs and then to observe the change of MHCC97-H cell invasion ability as well as the interventional effect of BMSCs on the animal model of hepatocellular carcinoma tissue MHCC97-H by fluorescence imaging in vivo. METHODS: MHCC97-H cells were divided into four groups: MHCC97-H group was set as a blank control group, and MHCC97-H NC siRNA as negative control group, and MHCC97-H siRNA TGFβ1 and siRNA OPN were experimental groups. Transwells assay was carried out for co-culture experiments. After 48 hours of co-culture, crystal violet staining was performed for cell counting in three randomly selected fields of vision. Combined with the red fluorescence protein gene, MHCC97-H cell lines in each group were inoculated via the right subaxillary subcutaneous transplantation to make a tumor model in nude mice. When the tumor volume was up to about 50 mm3, BMSCs were injected into the tumor in the nude mice, and 4 weeks later, fluorescence images were analyzed using software for fluorescence intensity. Frozen hepatocellular carcinoma tissue sections were taken for 4’,6-diamidino-2-phenylindole staining and fluorescence microscope observation. RESULTS AND CONCLUSION: Cell counting results showed that BMSCs significantly decreased MHCC97-H cells after gene silencing, and crystal violet staining showed that the migration ability of MHCC97-H cells was significantly decreased. Tumor volume shown by the fluorescence imaging was significantly reduced after the OPN gene transfection, the fluorescence intensity was lower than that in the other groups, and quantitative results showed that the absorbance value of OPN shRNA cells decreased significantly compared with other groups, indicating the BMSCs exhibit best interventional effectiveness in OPN-silenced MHCC97-H cells. Pathological sections showed that BMSCs were mainly distributed in the tumor necrosis area, and the fluorescence expression in the OPN siRNA group was more than that in the TGFβ1 siRNA group and the blank control group, indicating that after OPN gene silencing of MHCC97-H cells, the distribution of BMSCs in the tumor was increased. To conclude, it is able to reduce the invasive ability of hepatocellular carcinoma cells by inhibiting the expression of OPN and TGF β1 factors, and OPN silencing may be more conductive to BMSCs biotherapy.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of human umbilical cord versus placenta mesenchymal stem cells in prevention of mouse acute graft versus host disease Li Juan-juan, Wang You-wei, Ma Feng-xia, Du Wen-jing, Song Bao-quan, Wang Xin, Feng Ying, Tian Jian-jian,Han Zhong-chao 2017, 21 (5):  693-700.  doi: 10.3969/j.issn.2095-4344.2017.05.007 Abstract ( 452 )   PDF (8399KB) ( 273 )   Save BACKGROUND: Recently, the effects of human umbilical cord mesenchymal stem cells (hUCMSCs) and placenta-derived mesenchymal stem cells (PDMSCs) on treatment of acute graft versus host disease (aGVHD) have been confirmed in some in vitro studies or animal models. But there are still no reports comparing the therapeutic effects of these two cell types. OBJECTIVE: To compare the immunosuppressive function of hUCMSCs and PDMSCs in vitro or in a mouse aGVHD model. METHODS: (1) In vitro experiment. Human peripheral blood mononuclear cells (PBMCs) were isolated and divided into four groups: PBMCs cultured alone, PBMCs stimulated with phytohaemagglutinin (PHA), PHA stimulated-PBMCs cocultured with hUCMSCs, PHA stimulated-PBMCs cocultured with PDMSCs. After 5 days, PBMCs proliferation and interferon-γ level in cell supernatant were measured. (2) In vivo experiment. Fifty-seven BABL/C(H-2d) mice exposed to 8.5 Gy irradiation were randomly divided into five groups: only saline injection group, syngeneic bone marrow transplantation group, allogeneic bone marrow transplantation group, aGVHD group, hUCMSCs treatment group, PDMSCs treatment group. The clinical aGVHD score, histopathology of skin, liver, and small intestine, and survival time were analyzed at days 11, 14, 21 after transplantation. RESULTS AND CONCLUSION: (1) In vitro test: compared with the hUCMSCs, PDMSCs had stronger anti-inflammatory function. (2) In vivo test: The clinical scores on acute graft versus host disease were significantly lower in the hUCMSCs and PDMSCs treatment groups than that in the aGVHD group (P < 0.05). The survival rates of mice were significantly increased in the hUCMSCs and PDMSCs treatment groups compared to the aGVHD group (P < 0.05). Evident skin lesions were not found in all groups. Although small intestine mucosal lesions were found in all groups, the damage level seemed similar. Notably, significant difference was found in the liver that multifocal necrosis and a large number of inflammatory cells were seen in the aGVHD group, but less necrosis and inflammatory cells in the hUCMSC and PDMSC treatment groups. In conclusion, hUCMSC and PDMSC are comparably effective in the treatment of aGVHD in mice.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of dental pulp stem cell transplantation on the long-term behavior and cAMP response element binding protein in neonatal rats with hypoxic ischemic brain damage Wang Ai, Mu Qing-jie, Wang Xiao-li, Yan Shao-zhen, Qu Peng-yu, Wang Hai-yu, Hu Wen-ting 2017, 21 (5):  701-706.  doi: 10.3969/j.issn.2095-4344.2017.05.008 Abstract ( 313 )   PDF (1669KB) ( 174 )   Save BACKGROUND: cAMP response element binding protein (CREB) is a key protein of memory, which is closely related to long-term memory. It will provide a new way for the treatment of hypoxic ischemic brain damage (HIBD) to study the effects of dental pulp stem cells transplantation on the long-term behavior and CREB protein via the lateral ventricle in neonatal HIBD rats. OBJECTIVE: To observe the changes in long-term behavior and CREB protein expression in neonatal HIBD rats after human dental pulp stem cell transplantation, thereby providing scientific evidence for clinical treatment of neonatal HIBD.  METHODS: Thirty-six healthy 7-day-old Sprague-Dawley rats were randomly divided into normal, HIBD and cell transplantation group. The hypoxic ischemic brain damage models were established in the brain damage and cell transplantation groups. Twenty-four hours after HIBD, human dental pulp stem cells were injected into the left lateral cerebral ventricle of rats in the cell transplantation group, totally 3×106 living cells. Equal volume of normal saline was injected into the left lateral cerebral ventricle of rats in the normal control and HIBD groups. RESULTS AND CONCLUSION: The average time to seek water, the average escape latency and escape distance of the human dental pulp stem cells group were significantly shorter than those of hypoxic ischemic brain injury group (P < 0.01), but longer than those in the normal group (P < 0.01). Nissl staining showed that the cells in the hippocampal CA1 region in human dental pulp stem cells group were more regular, the number of cells was significantly higher than that of hypoxic ischemic brain injury group, but still significantly less than that in the normal group (P < 0.05). Immunohistochemical staining results showed that the number of CREB positive cells in human dental pulp stem cells group was significantly higher than those in HIBD group, but still significantly less than those in the normal group (P < 0.01). It is suggested that human dental pulp stem cells transplantation could promote the expression of CREB protein in the hippocampal CA1 region, to improve the long-term learning and memory ability of hypoxic ischemic neonatal rats, and thus repair HIBD.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of human amniotic mesenchymal stem cells transplantation via tail vein on neurological function recovery from ischemic reperfusion injury Zhao Yu 2017, 21 (5):  707-712.  doi: 10.3969/j.issn.2095-4344.2017.05.009 Abstract ( 292 )   PDF (1114KB) ( 180 )   Save BACKGROUND: Studies have shown that overexpression of myelin-associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMgp) is the main reason for early neuronal regeneration failure. OBJECTIVE: To study the effect of human amniotic mesenchymal stem cells (hAMSCs) transplantation on neural functional recovery of rats with ischemia/reperfusion (I/R) injury. METHODS: Sixty Sprague-Dawley were randomized into sham, I/R, and hAMSCs groups (n=20 per group). Rats in the hAMSCs group were given 1 mL of hAMSCs suspension (1.0×108/L) via the tail vein 24 hours after I/R injury. Rat neurological function recovery was assessed based on behavior changes, as determined by Longa behavioral score, cylinder test, horizontal ladder walking test and limb symmetry test at 1, 3 days, and 1, 2, 3 weeks post transplantation. Cell migration and distribution were observed using immunofluorescence method at 1 day, and 1, 2, 3 weeks post transplantation. MAG and OMgp protein expression was detected by western blot assay at 2 weeks post transplantation. Neuronal apoptosis was detected by TUNEL staining at 3 weeks post transplantation. RESULTS AND CONCLUSION: In the hAMSCs transplantation group, red marker-positive cells were visible around the injury region at 1 week after transplantation, and over time, these cells were increased in number. Significant improvement in the neurological function of rats were observed in the hAMSCs group as compared with the I/R group at 3 days and 1, 2, 3 weeks after transplantation (P < 0.05), and the expression of MAG and OMgp proteins were also decreased dramatically in the hAMSCs group (P < 0.05). After I/R injury, the number of apoptotic cells was increased, but hAMSCs transplantation reversed this effect. Overall, hAMSCs transplantation can reduce neuronal apoptosis by reducing MAG and OMgp expression levels, and thereby promote neurological functional recovery from I/R injury in rats.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of magnetic labeled endothelial progenitor cell transplantation on renal function of diabetic rats: a MRI imaging verification Feng Na, Xu Ying-jin, Dong Xi, Yang Jing-jing, He Xin 2017, 21 (5):  713-717.  doi: 10.3969/j.issn.2095-4344.2017.05.010 Abstract ( 263 )   PDF (983KB) ( 167 )   Save BACKGROUND: Endothelial progenitor cells have been shown to play an important role in the pathogenesis of traumatic diseases in recent years. OBJECTIVE: To explore the effect of magnetic labeled endothelial progenitor cell transplantation on renal  function of diabetic rats through a MRI imaging study. METHODS: Sixty Wistar rats were randomly divided into normal (no treatment), control and experimental groups. Intraperitoneal injection of 40 mg/kg streptozotocin was performed to make a rat model of type 1 diabetes in the control and experimental groups. Four weeks after modeling, rats in the experimental group were given intravenous injection of magnetic labeled endothelial progenitor cells (0.15 mL, 1×109/L). Fasting blood glucose, serum insulin, serum creatinine, urea nitrogen and 24-hour urinary protein levels in rats were measured at 8 weeks after cell transplantation. MRI was used to trace transplanted cells in vivo in comparison with renal biopsy findings, and rat body mass and kidney weight were measured to calculate kidney weight index. RESULTS AND CONCLUSION: After modeling, fasting blood glucose, serum creatinine, urea nitrogen and 24-hour urinary protein levels as well as kidney weight index were increased significantly (P < 0.05), while the insulin level decreased (P < 0.05). Compared with the model group, the endothelial progenitor cell transplantation reversed these indices (P < 0.05). Additionally, in the experimental group, there was slightly longer T1 and shorter T2 signals as well as marked lesion edge, and the FLASH sequence became more remarkable compared with the T2-weighted RARE sequence. The other groups showed no significant low signal changes. Magnetic-labeled positive cells in the experimental group showed by the MRI were consistent with the tissue biopsy results, while no positive cells were found in the model and normal groups. To conclude, the magnetic labeled endothelial progenitor cell transplantation can improve renal dysfunction in diabetic rats to a certain extent.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Co-culture with vascular endothelial progenitor cells: effects on proliferation and apoptosis of neural stem cells and vascular remodeling in rats with ischemia reperfusion injury Yang Chun-sheng, He Dan, Tan Jun 2017, 21 (5):  718-723.  doi: 10.3969/j.issn.2095-4344.2017.05.011 Abstract ( 300 )   PDF (4442KB) ( 207 )   Save BACKGROUND: Neural stem cell (NSC) transplantation is a common method for various ischemic  encephalopathies, but inability to survive in the transplantation region limits its further use in clinical practice. OBJECTIVE: To explore the effect of vascular endothelial progenitor cells (VEPCs) on the proliferation and apoptosis of co-cultured NSCs as well as vascular remodeling in rats with ischemia reperfusion injury. METHODS: 125 Sprague-Dawley rats were randomly divided into five groups, 25 rats in each group, including sham operation, ischemia, NSCs, co-culture, and VEPCs groups. Rat models of ischemia reperfusion injury were made in all groups except for the sham operation group, followed by corresponding interventions. The proliferation and apoptosis of neural stem cells were detected, and vascular remolding in the ischemic region was observed in each group. RESULTS AND CONCLUSION: At different time points after transplantation, BrdU positive cells were not observed in VEPCs, ischemia and sham operation groups; the number of BrdU positive cells in the co-culture group was significantly higher than that in the NSCs group (P < 0.05); BrdU+/Caspase-3+ cell were observed in both co-culture and NSCs groups, and the apoptosis rate of the co-culture group was significantly lower than that in the NSCs group (P < 0.05); there were new blood vessels in all the groups except for the sham operation group, and the number of new bone vessels was highest in the co-culture group. To conclude, our experimental results show that VEPCs promotes the proliferation of co-cultured NSCs, inhibits cell apoptosis and and promote angiogenesis in the ischemic penumbra of rats with ischemia reperfusion injury.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of Buyang Huanwu decoction combined with bone marrow mesenchymal stem cell transplantation on expression of integrin in a rat model of middle cerebral artery occlusion Zhang Yun-ke, Yang Jun-hong, Gao Feng, Duan Feng-yang 2017, 21 (5):  724-729.  doi: 10.3969/j.issn.2095-4344.2017.05.012 Abstract ( 332 )   PDF (5948KB) ( 263 )   Save BACKGROUND: Previous studies have found that combined use of Buyang Huanwu decoction and bone marrow mesenchymal stem cell (BMSC) transplantation can play a synergic role against cerebral ischemia injury. OBJECTIVE: To analyze the effect of Buyang Huanwu decoction combined with BMSC transplantation to promote angiogenesis after cerebral ischemia. METHODS: Ninety-six Sprague-Dawley rats were randomly divided into four groups, and used to make middle cerebral artery occlusion models. In combined group, rats were given intragastrical administration of Buyang Huanwu decoction 10 mL/kg once a day, beginning at 3 days prior to modeling, and then given intragastrical administration of Buyang Huanwu decoction once at 2 hours after modeling, followed by its intragastrical administration every 12 hours. In BMSC and combined groups, BMSC suspension was injected into the rat ventricle after 2-hour cerebral ischemia/2-hour reperfusion, and then 30 minutes later, CD34 and CD45 antibodies were injected. In antibody group, CD34 and CD45 antibodies were injected. In model group, only normal saline was given. Serum αVβ3 level detection, immunohistochemical observation, Q-PCR and western blot tests were performed in the combined group at 12, 24, 36, 48 hours after reperfusion, while these indices were detected in the other three groups at 36 hours after reperfusion. RESULTS AND CONCLUSION: (1) The level of serum αVβ3 was lower in the antibody group than the model group (P < 0.05), higher in the BMSC and combined groups than the antibody group (P < 0.05), and higher in the combined group than the BMSC group (P < 0.05). (2) Immunohistochemical findings showed that compared with the antibody group, the number of CD34 positive cells was higher in the model, BMSC and combined groups (P < 0.05). (3) Results from the Q-PCR and western blot assay showed that compared with the model group, the pFAK protein expression level was lower in the antibody and BMSC group, but FAK gene expression level had no overt changes; while the protein levels of FAK (24 hours after reperfusion) and pFAK (12 hours after reperfusion) were significantly increased in the combined group than the antibody and BMSC groups (P < 0.05,P < 0.01). Moreover, this increase exhibited a gradually rising trend with the extension of reperfusion time. To conclude, the combined use of BMSC transplantation and Buyang Huanwu decoction can reverse the effect of CD34+CD45 antibodies that lead to the decrease in the number of vascular endothelial cells and levels of integrin αVβ3 and downstream signaling molecules, thereby to promote angiogenesis in the MCAO model.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus Wang Rong-guo, Pi Li-hong, Chen Hong-yao, Zhang Hai-zhong 2017, 21 (5):  730-735.  doi: 10.3969/j.issn.2095-4344.2017.05.013 Abstract ( 326 )   PDF (5774KB) ( 282 )   Save BACKGROUND: Neural stem cells have multi-directional differentiation potential, self-sustaining and self-renewal capacity as well as have strong migration ability. Bushen Huoxue Recipe can reduce neuronal damage and promote nerve cell regeneration, to achieve neural function reconstruction. Underlying mechanisms of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus induced by sodium salicylate are yet unclear. OBJECTIVE: To investigate the effects of Bushen Huoxue Recipe combined with neural stem cell transplantation in rats with tinnitus induced by sodium salicylate. METHODS: Sixty Sprague-Dawley rats were randomized into five groups (n=12): normal control group, tinnitus model group, Bushen Huoxue Recipe group, stem cell group and combined treatment group (Bushen Huoxue Recipe combined with neural stem cell transplantation). Animal models of tinnitus induced by sodium salicylate were made in all the groups except for the normal control group. Fifteen days after modeling, rats were given intragastric administration of Bushen Huoxue Recipe water decoction (3 mL, 2.592 g/mL) for consecutive 7 days in the Bushen Huoxue Recipe group, intravenous injection of neural stem cells (1 mL, 1.0×109/L) in the stem cell group, or their combined treatment in the combined treatment group. RESULTS AND CONCLUSION: Bushen Huoxue Recipe, neural stem cell transplantation and their combination all could effectively promote the recovery of drinking water inhibitory rate that was ranked as follows: combined treatment group < Bushen Huoxue Recipe group < stem cell group. Additionally, the above-mentioned methods could effectively reduce pathological damage of primary auditory cortex in the rat brain. Western blot results showed higher 5-serotonin levels in the Bushen Huoxue Recipe group and combined treatment group than in the stem cell group, which were identical to ELISA findings. Our experimental findings indicate that Bushen Huoxue Recipe combined with neural stem cell transplantation exhibit certain effects on tinnitus induced by sodium salicylate in rats, and the mechanism of action is associated with neuronal repair and expression of 5-serotonin at mRNA and protein levels.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Intra-articular injection of Sox9-transfected bone marrow mesenchymal stem cells for treatment of knee osteoarthritis Zhuo Qun-hao, Zhang Wei-na, Li Jian, Wang Hong-wei, Zheng Feng, Ying Bin-bin 2017, 21 (5):  736-741.  doi: 10.3969/j.issn.2095-4344.2017.05.014 Abstract ( 331 )   PDF (4347KB) ( 192 )   Save BACKGROUND: Bone marrow mesenchymal stem cells are considered to have good proliferation and differentiation potentials. Sox9 is a transcription factor that is essential for chondrogenesis and has been termed as a “master regulator” of the chondrocyte phenotype. OBJECTIVE: To study the therapeutic effects of Sox9-transfected bone marrow mesenchymal stem cells on knee osteoarthritis. METHODS: The bone marrow mesenchymal stem cells were transfected with Lenti-Sox9-EGFP in vitro. The model of murine knee osteoarthritis was established by cutting off the anterior cruciate ligament. Thirty model mice were randomly divided into three groups, as normal saline group, bone mesenchymal stem cell group and Sox9-transfected bone mesenchymal stem cell group. 0.1 mL of normal saline, 0.1 mL of normal saline containing non-transfected bone marrow mesenchymal cells (non-transfected group), or 0.1 mL of normal saline containing Sox9-transfected bone marrow mesenchymal cells (Sox9-transfected group) was injected into the knee joint cavity of mice in the corresponding group, respectively. After 4, 8, 12 weeks, the repair of articular cartilage lesions was evaluated by toluidine blue and immunohistochemical staining. RESULTS AND CONCLUSION: The lesions of articular cartilage were more serious in the normal saline group, compared with the other two groups, and the difference became more obvious over time. Damaged articular cartilage was improved in the non-transfected group, but the improvement was less than that in the Sox9-transfected group. Immunohistochemistry staining revealed that in the Sox9-transfected group, the positive type II collagen expression was stronger than that in the other two groups, but this positive expression was decreased over time in all the three groups. These results suggest that Sox9-transfected bone marrow mesenchymal stem cells promote the repair of damaged cartilage in mice with knee osteoarthritis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of mesenchymal stem cells on airway inflammation in asthmatic mice depleted of CD4+CD25+ regulatory T cells Zhuansun Yong-xun, Zhang Wei, Du Yu-mo, Ran Pi-xin, Chen Rui, Lin Lin, Li Jian-guo 2017, 21 (5):  742-747.  doi: 10.3969/j.issn.2095-4344.2017.05.015 Abstract ( 261 )   PDF (4967KB) ( 218 )   Save BACKGROUND: Mesenchymal stem cells have an immunoregulatory capacity to suppress the airway inflammation of asthmatic mice, but the mechanism remains unclear. Our preliminary studies indicated that mesenchymal stem cells can upregulate the CD4+CD25+ regulatory T cells (Treg) of the asthmatic mice. OBJECTIVE: To study the effect of mesenchymal stem cells on the airway inflammation in asthmatic BALB/c mice depleted of CD4+CD25+ Treg. METHODS: Forty BALB/c mice were randomly divided into five groups: normal control group (group A), asthmatic group (group B), asthmatic group depleted of CD4+CD25+Treg (group C), mesenchymal stem cells group (group D), and asthmatic group depleted of CD4+CD25+Treg and administrated with mesenchymal stem cells (group E). Except group A, mice in the other groups were sensitized and challenged by ovalbumin to establish asthma models. In group C and group E, each mouse was treated with anti-CD25+ monoclonal antibody to deplete CD4+CD25+Treg. In group D and group E, mice were intravenously administered with 0.2 mL mesenchymal stem cells 1×109/L) at 10 days after sensitization. At 24 hours after the final activation, the number of CD4+CD25+Treg in the peripheral blood was detected by flow cytometry. The total cell number of inflammatory cells in the bronchoalveolar lavage fluid, and the number of eosinophils, lymphocytes and neutrophils were counted to analyze the degree of inflammation of the airway together with pathological observation. RESULTS AND CONCLUSION: (1) The proportion of CD4+CD25+Treg in peripheral lymphocytes was ranged as follows: group B < group D < group A (P < 0.01). (2) Number of inflammatory cells in the bronchoalveolar lavage fluid: Regarding the total cell number and the number of eosinophils in the bronchoalveolar lavage fluid, ranking was as follows: groups B, C, D, E were significantly higher than group A (P < 0.01), group C was significantly higher than group B ( P < 0.01), group D was significantly lower than group B (P < 0.01), group E was significantly lower than group C (P < 0.01). (3) Pathological observation of the lung: Mesenchymal stem cells could significantly inhibit the airway inflammation of asthmatic mice. The asthmatic mice appeared to have severer airway inflammation when CD4+CD25+Treg were depleted from the peripheral blood. However, mesenchymal stem cells could also significantly inhibit the airway inflammation of asthmatic mice depleted of CD4+CD25+Treg. Taken together, our data indicate that mesenchymal stem cells can significantly inhibit the airway inflammation of asthmatic mice depleted of CD4+CD25+Treg.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Chemokine (C-X-C motif) ligand 8 enhances the homing ability of human umbilical vein endothelial cells by promoting a paracrine response in mesenchymal stem cells under the high glucose environment Xie Li-ping, Zhang Shan-qiang, Sun Shi-zhu, Zhang Hai-yan, Lang Wei-ya, Zhang Meng, Shen Lei 2017, 21 (5):  748-754.  doi: 10.3969/j.issn.2095-4344.2017.05.016 Abstract ( 330 )   PDF (5722KB) ( 215 )   Save BACKGROUND: Chemokines can promote (MSCs) the secretion of vasoactive factors from mesenchymal stem cells (MSCs) through paracrine mechanism, which have important role in accelerating angiogenesis. OBJECTIVE: Under the high glucose environment, to the effect of the supernatant of MSCs stimulated by chemokine (C-X-C motif) ligand 8 (CXCL-8) on human umbilical vein endothelial cells (HUVECs), and to analyze the mechanism of Sonic Hedgehog signaling pathway in the stimulation of CXCL-8 on MSCs. METHODS: Under the high glucose environment, the MSCs supplemented with 100 μg/L CXCL-8 were set as CXCL-8 group; the MSCs that were preprocessed with 5 μmol/L octyl maleimide for 45 minutes and then stimulated with 100 μg/L CXCL-8 were as Shh inhibitor group; the MSCs that were routinely cultured in a high-glucose medium were as control group. The cell supernatant of each group was extracted as conditioned medium (CM) to culture HUVECs, respectively, and these cells were referred to as CXCL-8 CM group, Shh inhibitor CM group, and control CM group, respectively. Cell counting kit-8, cell scratch and Transwell chamber tests were used to observe the effect of each CM on HUVEC proliferation, apoptosis and chemotaxis. By establishment of a diabetic skin ulcer model in C57BL/6J mice, the CM of each group was used to treat the mouse model to confirm the effects of CXCL-8 stimulated MSCs CM on HUVEC homing and ulcer healing. RESULTS AND CONCLUSION: (1) The experimental results in vitro: compared with the control CM group, CXCL-8 CM group significantly promoted the proliferation of HUVECs, and decreased the apoptosis of HUVECs, the closure rate and migration rate of HUVECs were significantly increased (P < 0.01 or P < 0.01), and the levels of vascular endothelial growth factor and epidermal growth factor were significantly increased (P < 0.01 or P < 0.01). Compared with CXCL-8 CM group, however, the above results in the Shh inhibitor CM group showed reverse changes (P < 0.01). (2) The experimental results in vivo: compared with the MSCs CM group and Shh inhibitor CM group, the healing effect of diabetic skin ulcer and the number of HUVECs labeled by green fluorescent protein in the CXCL-8 CM group were significantly increased (P < 0.01). To conclude, these findings indicate that CXCL-8 stimulated MSCs secrete paracrine factors, vascular endothelial growth factor and epidermal growth factor, through the Sonic Hedgehog signaling pathway under the high glucose environment, which enhance the homing ability of HUVECs.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biological characterization and in vitro biocompatibility of human placenta derived mesenchymal stem cells Wu Jia, Wen Yong-mei, Lv Xin-rong, Mu Yan-dong 2017, 21 (5):  755-759.  doi: 10.3969/j.issn.2095-4344.2017.05.017 Abstract ( 283 )   PDF (2246KB) ( 206 )   Save BACKGROUND: At present bone marrow mesenchymal stem cells act as the main seed cells in bone tissue engineering, but only 0.001%-0.01% cells are in the bone with difficulty in cell separation and purification. OBJECTIVE: To explore the biological characterization of human placenta derived mesenchymal stem cells and  biocompatibility with three-dimensional porous hydroxyapatite ceramic scaffold. METHODS: Human placenta derived mesenchymal stem cells were morphologically observed and identified using flow cytometry, followed by osteogenic, adipogenic, chondrogenic induction for 3 weeks. Afterwards, the potential of multi-directional differentiation was identified by alizarin red S, oil red O and toluidine blue staining. DAPI staining was used to observe the adhesion of cells on the surface of the hydroxyapatite ceramic scaffold under scanning electron microscope. RESULTS AND CONCLUSION: The human placenta derived mesenchymal stem cells showed long spindle shape and uniform size under the microscope; they highly expressed CD29 and D90, but did not express CD45 and CD106. Following induction, mineralized nodules were observed by alizarin red S staining, lipid droplets by oil red O staining and blue-dyed toluidine blue staining. These cells adhered well to the scaffold surface, indicting they are suitable for bone tissue engineering.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Endometrial stromal stem cells: in vitro isolation, culture and biological features Bi Yu-hu, Teng Jun-ru 2017, 21 (5):  760-765.  doi: 10.3969/j.issn.2095-4344.2017.05.018 Abstract ( 326 )   PDF (4786KB) ( 203 )   Save BACKGROUND: Understanding and studying endometrial cells from the perspective of stem cells can provide a new therapeutic approach and research entry point for the clinical treatment of endometrial diseases. OBJECTIVE: To compare the biological features of endometrial stromal stem cells isolated and cultured using different methods. METHODS: Three commonly used cell separation methods, including trypsin, collagenase I and their combination, were used to isolate and culture endometrial stromal stem cells from the uterus after removal. Afterwards, passage 2 cells cultured for 5 days were subjected to trypan blue staining and living cell counting. Immunohistochemical staining for CD146 and CD90 and RT-PCR were used to identify harvested endometrial stromal stem cells. Logarithmically growing cells were cultured to draw cell growth curves. RESULTS AND CONCLUSION: Harvested endometrial stromal stem cells presented with spindle, adherent cell growth, nonpolar arrangement, and fibroblast morphology. The DNA of endometrial stem cell marker CD146 and CD90 highly expressed in 500 bp and 150 bp respectively, but did not express in the mesometrium. S-shaped growth curve of cultured cells was found, and there was no difference in the cell cycle of cells cultured using three different methods. Among the three methods, collagenase I method could harvest the highest number of endometrial stromal stem cells that grew fastest. These findings indicate that endometrial stromal stem cells can be successfully isolated from the removed uterine tissues, and collagenase I method is an efficient, practical and stable method for in vitro culture of endometrial stromal stem cells.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of long non-coding RNA NR_033474 on proliferation of C3H10T1/2 mesenchymal stem cells Pan Ya-qiong, Dai Zhong, Zuo Chang-qing, Wang Zong-gui 2017, 21 (5):  766-772.  doi: 10.3969/j.issn.2095-4344.2017.05.019 Abstract ( 242 )   PDF (1206KB) ( 283 )   Save BACKGROUND: Recent studies have found that long non-coding RNAs (lncRNAs) can regulate stem cell proliferation and differentiation. But it is unclear that how lncRNA NR_033474 regulate stem cell proliferation and cell cycle. OBJECTIVE: To investigate the effect of lncRNA NR_033474 on the proliferation and cell cycle regulation in C3H10T1/2 mesenchymal stem cells after the NR_033474 overexpressed by lentivirus, and to study the possible regulation mechanism of NR_033474 on mesenchymal stem cells. METHODS: LncRNA NR_033474 was cloned into a lentivirus vector. Lentivirus particles were infected into C3H10T1/2 cells to upregulate the expression of NR_033474. The NR_033474 expression level was detected by real-time PCR. Compared with the empty lentivirus vector, the proliferation of C3H10T1/2 cells which overexpressed NR_033474 was detected by cell counting assay and cell cycle was detected using flow cytometry. The expression of cell cycle-associated proteins such as CDK1, Cyclin B1, Cyclin D1 and P53 were detected by western blot assay. RESULTS AND CONCLUSION: Compared with the control group, lncRNA NR_033474 in C3H10T1/2 cells which overexpressed NR_033474 was increased by about 100 times (P < 0.01), and the proliferation of C3H10T1/2 cells was significantly inhibited after NR_033474 overexpression by lentivirus (P < 0.05). In addition, flow cytometry showed that C3H10T1/2 cells overexpressing NR_033474 were arrested in G2/M phase compared to the control group. Western blot showed that the expression levels of CDK1 and Cyclin B1 were downregulated, while there were no changes in Cyclin D1 and P53 expression. To conclude, these findings suggest that the NR_033474 overexpression significantly inhibits the cell growth of C3H10T1/2 cells, at least in part, through induction of cell cycle arrest at G2/M phase.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Anti-oxidative activities of supernatants of human fetal placental mesenchymal stem cells cultured in serum-free medium Fu Xue, Zhang Yu-jie, Yan Xiu-rui, Ma Xiao-na, Liu Xiao-ming, Wei Jun 2017, 21 (5):  773-779.  doi: 10.3969/j.issn.2095-4344.2017.05.020 Abstract ( 322 )   PDF (1176KB) ( 286 )   Save BACKGROUND: Current research on mesenchymal stem cells (MSCs) is mostly focused on its immune  regulatory function, while little is reported on the antioxidant capacity of the cells and culture supernatant. OBJECTIVE: To investigate the anti-oxidative capacity of the supernatant harvested from human fetal placenta MSCs (fPMSCs) under a condition of serum free culture. METHODS: fPMSCs were cultured with serum free media, and the supernatants of cells at passages 2-6 were collected at 48 hours after culture. Vitamin C was added into the culture medium, as a positive control, and its concentration was 100 μmol/L. The total antioxidant capacity, scavenging capacity of free radicals and antioxidant enzymatic activities of supernatants were measured. RESULTS AND CONCLUSION: By comparing anti-oxidative activities of vitamin C and naïve culture medium, supernatants collected from fPMSCs cultures exhibited obvious antioxidant capacities at different extents between passages of cell cultures. The total antioxidant capacity of the culture supernatant was comparable to 40-80 μmol/L vitamin C. In addition, all supernatants derived from cells with different passages displayed capacities to scavenge free radicals, including 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•), hydroxyl radical (•OH), superoxide anion radical (O2-). Even more, activities of antioxidant enzymes, including superoxide dismutase and glutathione peroxidase, were also detected in supernatants collected from different passages of fPMSCs. Under the serum-free condition, the culture supernatants of fPMSCs have antioxidant capacities at certain extent. However, the antioxidant components and underlying mechanisms need to be further studied.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Safety and efficacy of autologous bone marrow mesenchymal stem cells for dilated cardiomyopathy: a Meta-analysis Ai Jin-wei, Liu Ying, Liu Chu-fan, Pei Bin 2017, 21 (5):  780-788.  doi: 10.3969/j.issn.2095-4344.2017.05.021 Abstract ( 401 )   PDF (1546KB) ( 254 )   Save BACKGROUND: Autologous bone marrow mesenchymal stem cells (BMSCs) transplantation has been used for clinical treatment of dilated cardiomyopathy. But the efficacy and safety of autologous BMSCs transplantation remains controversial. OBJECTIVE: To systematically assess the efficacy and safety of autologous BMSCs transplantation for treatment of dilated cardiomyopathy by using meta-analysis approach. METHODS: PubMed, Cochrane Library (Issue 2, 2016), Embase, CNKI, CBM, VIP, WanFang were systemically searched for relevant randomized controlled trials (RCTs) about autologous BMSCs transplantation and conventional drugs for the treatment of dilated cardiomyopathy. After information extracting and quality assessing, Meta-analysis of left ventricular ejection fraction, left ventricular end-diastolic diameter, 6-minute walking distance, percentage of myocardial perfusion defect, mortality, incidence of malignant arrhythmia events and heart transplantation rate during treatment and follow-up was performed using R3.1.0 software. RESULTS AND CONCLUSION: A total of 7 RCTs involving 341 patients were included. Meta-analysis results showed that for efficacy, compared with the conventional drugs, BMSCs can increase the left ventricular ejection fraction [1 month post-treatment: mean difference (MD)=3.02, 95% confidence interval (CI) (1.55, 4.49); 3 months post-treatment: MD=4.38, 95%CI(3.55, 5.52); 6 months post-treatment: MD=6.47, 95%CI(4.78, 8.15); ≥ 12 months post-treatment: MD=8.23, 95%CI(5.15, 9.19)]; decrease the left ventricular end-diastolic diameter after 3 months [3 months post-treatment: MD=-0.65, 95%CI(-0.72, -0.59); 6 months post-treatment: MD=-0.12, 95%CI(-0.21, -0.03); ≥ 12 months post-treatment: MD=-0.19, 95%CI(-0.24, -0.13)]; increase 6-minute walking distance after 6 months [6 months post-treatment: MD=87.70, 95%CI(51.55, 123.85); ≥ 12 months post-treatment: MD=143.83, 95%CI(122.73, 164.93)]; and decrease percentage of myocardial perfusion defect at 3 months [MD=-3.56, 95%CI(-5.57, -1.55)]. For safety, BMSCs can decrease the mortality [risk ratio=0.46, 95%CI(0.24, 0.89)], but there is no significant difference in the incidence of malignant arrhythmia events and heart transplantation rate between two treatment groups. To conclude, these results indicate that BMSCs transplantation for dilated cardiomyopathy is one of effective and safe treatments.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Research progress and application outlook of paracrine functions of adipose-derived stem cells in facial anti-aging Guo Ji-an, Yu Pi-jun, Wang Lu-ping, Shi Ying-ying, Liu Yi, Chen Wei 2017, 21 (5):  789-794.  doi: 10.3969/j.issn.2095-4344.2017.05.022 Abstract ( 426 )   PDF (903KB) ( 335 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs) are a subset of mesenchymal stem cells obtained from adipose tissue, which have paracrine functions, already becoming a focus in anti-aging researching. OBJECTIVE: To review the research progress and application outlook of ADSCs’ paracrine functions in facial anti-aging. METHODS: The first author searched the PubMed and CNKI databases using the keywords of “adipose-derived stem cells, paracrine, anti-aging” in English and Chinese, respectively, to retrieve relevant articles published from January 2001 to May 2016. Repetitive articles or those with no originality were eliminated. Totally 176 articles were searched initially, and 66 articles were included in result analysis. RESULTS AND CONCLUSION: ADSCs have paracrine function, which can secrete various growth factors (e.g., epidermal growth factor, transforming growth factor β, platelet-derived growth factor, vascular endothelial growth factor, collagen and fibronectin) and inflammatory factors (e.g., interferon-γ, interleukin-lβ, interleukin-8, interleukin-9, interleukin-12, interleukin-15, interleukin-17 and tumor necrosis factor-α). These paracrine products of ADSCs have significant effects on anti-aging, such as inhibiting skin aging, whitening skin, assisting lipotransfer, promoting hair regeneration.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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The biological features of adipose-derived stem cells and its application in oral tissue regeneration Luo Yuan, Huang Yuan-liang 2017, 21 (5):  795-801.  doi: 10.3969/j.issn.2095-4344.2017.05.023 Abstract ( 302 )   PDF (1101KB) ( 456 )   Save BACKGROUND: In comparison with stem cells from other tissues, adipose-derived stem cells (ADSCs) are not only abundant and easy to harvest but also have the strong abilities to proliferate and differentiate. These make ADSCS the promising stem cells in the field of oral tissue regeneration. OBJECTIVE: To summarize the biological features of ADSCs and the development of functions as stem cells in the oral tissue regeneration. METHODS: We took “adipose-derived stem cells, oral tissue regeneration, bone tissue engineering, periodontal tissue regeneration, bone defects” as the key words in Chinese and English, respectively, to retrieve the related literatures from PubMed, CNKI, Wangfang databases during 2000 to 2016 based on internet search. RESULTS AND CONCLUSION: In vitro studies on periodontal tissue regeneration have shown that ADSCs can promote the regeneration of bones, fibroblasts, blood vessels around the periodontal area, and improve the healing abilities of injured tissues. In the study of craniofacial bone regeneration, ADSCs can successfully repair massive bone loss due to the surgeries of tumor or accidents. ADSCs have a promising role in the oral tissue regeneration, but further studies will be required because of the problems of immunogenicity and safety.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells transplantation in the treatment of diabetic cystopathy Yang Ya-fei, Yang Jin, Chen Lin, Xing Sha-sha, Hu Hai-feng, Zhang Ya-mei, Wang Zi-li 2017, 21 (5):  802-808.  doi: 10.3969/j.issn.2095-4344.2017.05.024 Abstract ( 394 )   PDF (1037KB) ( 251 )   Save BACKGROUND: Stem cell transplantation has gained considerable support recently. It provides new opportunities for treating diabetic neurogenic bladder. OBJECTIVE: To summarize the research progress in bone marrow mesenchymal stem cells (BMSCs) transplantation in the treatment of diabetic neurogenic bladder. METHODS: The first author retrieved Sciencedirect, PubMed, Embase, Wangfang and CNKI databases, for relevant articles of BMSCs transplantation in the treatment of diabetic neurogenic bladder, published from 2000 to 2016. The key words were “bone marrow mesenchymal stem cells, diabetic neurogenic bladder, differentiation, transplantation” in Chinese and English, respectively. RESULTS AND CONCLUSION: In patients with diabetic neurogenic bladder, the transplantation of BMSCs may provide safer and longer-lasting outcomes by repairing the damaged bladder and urethra. And it can produce various bioactive substances, which will have nutritional paracrine effects on the bladder microenvironment, including anti-inflammation, promoting cell proliferation and improving cell survival. On the one hand, the BMSCs have the ability to migrate to the injury site via the blood circulation. On the other hand, BMSCs can produce various growth factors, as well as the cytokines that can inhibit the inflammatory response. While the current clinical studies are lacking, its efficacy and safety needs further verification.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Simvastatin regulates endogenous stem cells to reconstruct the degenerative intervertebral disc Huang Ze-nan, Feng Xin-min, Wang Jing-cheng, Chen Tao, Bi Song-chao, Zhang Liang 2017, 21 (5):  809-814.  doi: 10.3969/j.issn.2095-4344.2017.05.025 Abstract ( 353 )   PDF (938KB) ( 272 )   Save BACKGROUND: Statins can promote the mRNA expression of bone morphogenetic protein 2, aggrecan and type II collagen in intervertebral disc cells, and they also can reverse the phenotype of dedifferentiated nucleus pulposus cells to slow disc degeneration process. OBJECTIVE: To review the research progress of simvastatin modulating the biological characteristics and mobilizing endogenous stem cells for the repair of intervertebral disc degeneration.  METHODS: The first author retrieved the PubMed and CNKI databases for relevant articles published before January 2016 using the key words of “disc degeneration factor, Simvastatin AND stem cells, endogenous stem cells AND disc degeneration” in English and Chinese, respectively. Initially, 102 relevant articles were retrieved, but only 48 articles were included in result analysis following elimination of duplicate studies. RESULTS AND CONCLUSION: By summarizing a large number of studies on the treatment of intervertebral disc degeneration worldwide, we found that simvastatin may modulate the biological characteristics and function of nucleus pulposus mesenchymal stem cells via promoting the expression of hypoxia-inducible factor 1α for the endogenous stem cell-based therapy of intervertebral disc degeneration.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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The safety and application of induced pluripotent stem cells Tian Sheng-nan, Wang Bo, Li Qi, Huang Yuan-hua, Ma Yan-lin 2017, 21 (5):  815-820.  doi: 10.3969/j.issn.2095-4344.2017.05.026 Abstract ( 420 )   PDF (913KB) ( 207 )   Save BACKGROUND: Induced pluripotent stem cell technology have solved the contradiction between the ethics and immune rejection, and this high-efficient and safe technique is becoming the mainstream of today’s research. OBJECTIVE: To comprehensively review the safety and application of induced pluripotent stem cells. METHODS: A computer-based online retrieval of PubMed and CNKI was performed to search relevant papers published from January 2006 to April 2016, with the key words of “induced pluripotent stem cell, reprogramming, clinical application, safety, transcription factor, disease mode” in English and Chinese, respectively. RESULTS AND CONCLUSION: In recent years, research on induced pluripotent stem cells has attracted much attention from the scientific community and the medical community, and this technique has successfully gained induced pluripotent stem cells and overcome the problems of immunity and ethics. However, it is limited to the theoretical and laboratory research due to the inability to solve the safety, efficiency and re-differentiation mechanism of induced pluripotent stem cells. Therefore, we are faced with enormous difficulties and challenges, which involve all aspects of basic research, including how to safely and effectively induce the differentiation of induced pluripotent stem cells into the desired cell type and how to establish a suitable disease model as well as a high-throughput drug screening platform.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics