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08 January 2017, Volume 21 Issue 1 Previous Issue    Next Issue

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Establishment of a lentiviral vector carrying rat miR-21 gene and its effect on apoptosis of bone marrow mesenchymal stem cells Fu Xia-fei, He Yuan-li, Wang Xue-feng, Chen Xiao-ying 2017, 21 (1):  1-5.  doi: 10.3969/j.issn.2095-4344.2017.01.001 Abstract ( 416 )   PDF (931KB) ( 690 )   Save BACKGROUND: Apoptosis in bone marrow mesenchymal stem cells (BMSCs) occurs after transplantation, which is mainly responsible for unsatisfactory therapeutic effects on premature ovarian failure  induced by chemotherapy. miR-21 participates in the regulation of cell proliferation and apoptosis, and has important anti-apoptotic effect. OBJECTIVE: To construct a lentiviral vector for rat miR-21 and observe its effects on apoptosis of BMSCs induced by phosphoramide nitrogen mustard. METHODS: Rat miR-21 gene was synthesized, amplified, and connected with the lentiviral plasmid pLVX-shRNA2. Both double enzyme digestion and gene sequencing were done to identify the successful construction of the vector pLVX-shRNA2-rno-miR-21-5p. The vector was packaged, and the titer was examined. BMSCs were transfected by the vector with different multiplicities of infection (MOI=20 or 40), and the efficiency was observed. miR-21 expression in the transfected cells was detected using real-time PCR. Phosphoramide nitrogen mustard was added into the cell culture media of BMSCs, then the apoptotic rate of BMSCs was detected by flow cytometry, and apoptotic index was examined by Hoechst33342 staining. RESULTS AND CONCLUSION: The target gene was successfully connected with the lentiviral vector. The viral titer was 6×1011 CFU/L. The vector transfected BMSCs had a high efficiency above 90%. Real-time PCR results showed the expression levels of miR-21 in miR-21 group 1 (MOI=20) and miR-21 group2 (MOI=40) were higher than that in BMSCs group and blank vector group (P=0.000). Flow cytometry and Hoechst33342 staining results showed the apoptotic rate and apoptotic index of miR-21 transfected groups were lower than those of blank vector group and BMSCs group (P=0.001). Overall, we successfully established rat miR-21 lentivirus vector pLVX-shRNA2-rno-miR-21-5p and transfected it into the BMSCs. Upregulation of miR-21 could reduce BMSCs apoptosis, and enhance cell proliferation.   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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Influence of apelin on survival and vascularization potential of bone marrow mesenchymal stem cells under hypoxic and ischemic conditions Hou Jing-ying, Wang Lei, Zhong Ting-ting, Zhou Chang-qing, Guo Tian-zhu, Long Hui-bao, Wu Quan-hua, Zheng Shao-xin, Wu Hao, Wang Tong 2017, 21 (1):  6-12.  doi: 10.3969/j.issn.2095-4344.2017.01.002 Abstract ( 416 )   PDF (1228KB) ( 568 )   Save BACKGROUND: Our previous work demonstrated that bone marrow mesenchymal stem cells (BMSCs) transplantation could improve cardiac function in rats with myocardial infarction. However, the overall efficacy was not satisfactory. The implanted cells presented a low survival rate and newly formed vascular-like structures were sparse in the local infarct tissues. OBJECTIVE: To observe the influence of putative receptor protein related to the angiotensin receptor AT1 endogenous ligand (apelin) on the survival and vascularization potential of BMSCs in hypoxic and ischemic conditions and to investigate relevant mechanisms. METHODS: The bone marrow mesenchymal stem cells were obtained and cultured in vitro with or without apelin-13 (5 μmol/L) stimulation under hypoxic-ischemic condition (serum-free medium, 1% O2) for 24 hours, or cultured under normoxia (complete culture medium, 20% O2) as a negative control during the whole process. All the experimental groups were further co-cultured with human umbilical vein endothelial cells to promote vascular differentiation for another 6 hours. Cell proliferation, apoptosis, and vascular density were assessed and the expression of vascular endothelial growth factor was detected using western blot assay thereafter. RESULTS AND CONCLUSION: Compared with the BMSCs group, BMSCs+apelin group presented more rapid cell growth, higher proliferation rate, and lower apoptosis percentage under normoxic and hypoxic conditions. In the BMSCs+apelin group, a larger number of vascular branches formed on matrigel under normoxic and hypoxic-ischemic conditions; and the expression of vascular endothelial growth factor was significantly enhanced. These findings suggest that apelin could effectively promote BMSCs survival and vascularization under hypoxic-ischemic conditions in vitro. It might function via upregulating the expression of vascular endothelial growth factor.     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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Epithelial transdifferentiation of bone marrow mesenchymal stem cells promoted by ascorbic acid Han Xiao-lin, Li Jing-dong, Wang Wan-ling, Yang Cui, Li Zhi-ying 2017, 21 (1):  13-17.  doi: 10.3969/j.issn.2095-4344.2017.01.003 Abstract ( 464 )   PDF (1355KB) ( 547 )   Save BACKGROUND: Ascorbic acid has been shown to promote blood vessel elasticity, strengthen bones, and exert an anti-oxidative role. OBJECTIVE: To study the effect of ascorbic acid on the epithelial transdifferentiation of bone marrow mesenchymal stem cells in vitro by cell and molecular biology experiments. METHODS: Different concentrations (0, 10, 20, 40 μmol/L) of ascorbic acid were used to deal with bone marrow mesenchymal stem cells. Under light microscope, we observe the morphological changes of bone marrow mesenchymal stem cells, and screened the optimal concentration of ascorbic acid (40 μmol/L) and suitable interventional time (48 hours). Further detection of E-cadherin, N-cadherin, Snail1 and transforming growth factor-β by real-time PCR was conducted. Western blot assay was also used to detect Oct4, Nanog and  Sox2 in bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: Under the light microscope, the long-fusiform cells began to differentiate into rounded epithelial cells after 24 hours of ascorbic acid intervention. And this change became more obvious with the increasing of concentrations. After 48 hours, approximately 90% of cells changed as described above. Ascorbic acid upregulated N-, E- cadherin and downregulated Snail1 and transforming growth factor-β, thereby promoting the epithelial transition of bone marrow mesenchymal stem cells. Meanwhile, the expression of Sox2, Nanog, and Oct4, however, did not change significantly.   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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Differentiation of human umbilical cord mesenchymal stem cells into chondrocytes induced by direct co-culture with chondrocytes Li Xing-fu, Duan Li, Liang Yu-jie, Zhu Wei-min, Wang Da-ping 2017, 21 (1):  18-24.  doi: 10.3969/j.issn.2095-4344.2017.01.004 Abstract ( 333 )   PDF (4993KB) ( 789 )   Save BACKGROUND: Both chondrocytes and mesenchymal stem cells (MSCs) can be used to construct tissue-engineered cartilage. Chondrocytes cultured in vitro however are prone to dedifferentiation and difficult to maintain phenotypes, and accordingly, their clinical application is limited. OBJECTIVE: To explore the effect of human articular chondrocytes and human umbilical cord mesenchymal stem cells (hUC-MSCs) co-culture in vitro on the chondrogenic differentiation of hUC-MSCs and to optimize the co-culture ratio. METHODS: The hUC-MSCs surface marker was identified by flow cytometry. Human articular chondrocytes and hUC-MSCs were co-cultured at the ratio of 1:1, 3:1 and 5:1. The hUC-MSCs induced by transforming growth factor-beta 1 were set as a positive control group. Human articular chondrocytes and hUC-MSCs cultured alone were set as two negative control groups. The expression level of type II collagen (COL2) was analyzed by immunofluorescence staining. The protein expression of SRY-box9 (SOX9), type I collagen (COL1) and COL2 were determined by western blot. The mRNA levels of SOX9, Col1a1 and Col2a1 were detected by quantitative real-time PCR. RESULTS AND CONCLUSION: The hUC-MSCs were isolated from the human umbilical cord and identified with flow cytometry. After 28 days of culture, both the co-culture group and the positive control group were observed with positive staining under the immunofluorescence microscope. The Col2a1 mRNA expression level of the positive control group was higher than that of the co-culture group, but the total COL2 protein expression was lower. The Col2a1 mRNA expression level of the co-culture group in 1:1 was higher than that of the co-culture group in 3:1 or 5:1. Col1a1 mRNA and COL1 protein expression levels of the positive control group and co-culture group were lower than those of human articular chondrocyte negative control group. To conclude, the co-culture of hUC-MSCs and human articular chondrocytes significantly induces the hUC-MSC differentiation into chondrocytes and effectively restrains cell fibrosis. The optimal cell ratio in the co-culture system is demonstrated to be 5:1 (hUC-MSCs:chondrocytes). Therefore, the direct co-culture can be an economic way for inducing hUC-MSC differentiation into chondrocytes, which provides reliable seeding cells for cartilage 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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Transdifferentiation of human umbilical cord mesenchymal stem cells into cancer-associated mesenchymal stem cells induced by hepatocellular carcinoma HepG-2 supernatant Yang Juan, Zheng Sheng, Chen Wen-qin, Liu Jing, Zhang Fan, Wang Yu-bo 2017, 21 (1):  25-31.  doi: 10.3969/j.issn.2095-4344.2017.01.005 Abstract ( 343 )   PDF (5042KB) ( 342 )   Save BACKGROUND: Tumor microenvironment can recruit and attract different type and differently differentiated mesenchymal stem cells (MSCs) to the tumor growth site, thereby affecting tumor progression. OBJECTIVE: To investigate the transdifferentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cancer-associated MSCs induced by the supernatant from hepatocellular carcinoma cells HepG-2 culture medium and the effect of induced hUC-MSCs on the proliferation and migration of hepatocellular carcinoma cells HepG-2. METHODS: (1) Experiment 1: The supernatant from hepatocellular carcinoma cells HepG-2 culture medium was prepared, mixed with equal volume of low glucose DMEM and used to culture hUC-MSCs for 48 hours. The expressions of cancer-associated MSCs proteins and miR-221 in hUC-MSCs were detected. (2) Experiment 2: The supernatant from induced hUC-MSCs culture medium was mixed with equal volume of high glucose DMEM to treat hepatocellular carcinoma cells HepG-2 for 48 hours and then the proliferation and migration of hepatocellular carcinoma cells HepG-2 were observed. (3) Experiment 3: Hepatocellular carcinoma cells HepG-2 were cultured alone or co-cultured with hUC-MSCs for 48 hours and then the capabilities of proliferation and migration of hepatocellular carcinoma cells HepG-2 were observed. RESULTS AND CONCLUSION: (1) Experiment 1: The protein levels of vimentin and fibroblast activation protein were increased and the expression of miR-221 was upregulated in the hUC-MSCs after treated by the supernatant from hepatocellular carcinoma cells HepG-2 culture medium. (2) Experiment 2: The supernatant from induced hUC-MSCs culture medium promoted hepatocellular carcinoma cells HepG-2 proliferation and migration. (3) Experiment 3: The capabilities of proliferation and migration of hepatocellular carcinoma cells HepG-2 after co-cultured with hUC-MSCs were significantly enhanced. To conclude, these results above declared that the supernatant from hepatocellular carcinoma cells HepG-2 culture medium could induce hUC-MSCs, equipping them with cancer-associated MSC-like phenotype and promoting the proliferation and migration of hepatocellular carcinoma cells HepG-2.   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 tumorigenicity of bone marrow mesenchymal stem cells in the body after the conversion of gastric cancer microenvironment Wang Yong-feng, Liu Xi-ping, Cui Guo-ning, Dong Jun-gang, Li Pei-qing, Ming Hai-xia, Zhang Wei 2017, 21 (1):  32-37.  doi: 10.3969/j.issn.2095-4344.2017.01.006 Abstract ( 442 )   PDF (6247KB) ( 245 )   Save BACKGROUND: The clinical gastric cancer tissues and the mesenchymal stem cells isolated from tumor  tissues in the body of nude mice have similar characteristics with bone marrow mesenchymal stem cells, which are certificated to be an important part of the tumor microenvironment that can promote the growth of tumor. OBJECTIVE: To observe that whether bone marrow mesenchymal stem cells have the tumorigenic ability after the conversion of gastric microenvironment. METHODS: Bone marrow mesenchymal stem cells cultured singly and routinely were used as controls. Rat bone marrow mesenchymal stem cells were indirectly co-cultured with gastric cancer BCG-823 cells in a Transwell chamber to construct the gastric microenvironment (experimental group). Bone marrow mesenchymal stem cells from each group were subcutaneously seeded into the forelimb armpit of nude mice, and then, the formation of subcutaneous nodules was observed. Immunohistochemical method was used to detect the expression of tumor tissue latent membrane protein 1 (LMP1) and tumor suppressor gene TCF21. Transmission electron microscope was used to observe the ultrastructure of tumor tissues. RESULTS AND CONCLUSION: After 7-14 days, there were no tumor nodules in the nude mouse armpit in the control group. On the contrary, in the experimental group, tumor nodules formed in the nude mouse armpit after 7 days, and these nodules were enlarged and kept stable until the 14th day. The immunohistochemical results showed high level of LMP1, but low level of TCF21. Under the transmission electron microscope, the tumor tissues had a large volume and an irregular shape, the cell mricovilli were increased abnormally, and there were many fendritic protrusions on the surface, indicating the presence of obvious intracellular changes. In conclusion, bone marrow mesenchymal stem cells after the conversion in the gastric microenvironment have the ability of tumorigenicity in vivo.   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 human esophageal cancer mesenchymal stem cells on the proliferation and cell cycle of EC109 cells Qian You-hui, Peng Xue-feng, Wan Yan-hui, Gong Li-hong, Wang Wei, Gu Pei-gui 2017, 21 (1):  38-42.  doi: 10.3969/j.issn.2095-4344.2017.01.007 Abstract ( 424 )   PDF (3674KB) ( 284 )   Save BACKGROUND: Studies have shown that biological signals such as growth factors and cytokines released from tumor cells can make mesenchymal stem cells migrate into tumor tissues, and then these cells differentiate into various stromal cells constituting a microenvironment to participate in the regulation of tumor cells. OBJECTIVE: To investigate the effects of human esophageal cancer mesenchymal stem cells (hEC-MSCs) on the proliferation and cell cycle of human esophageal carcinoma cell line EC109. METHODS: After isolation and purification, hEC-MSCs at 1×104, 2×104, 3×104, 4×104 were co-cultured with  EC109 cells, respectively. MTT method and flow cytometry were used to measure cell proliferation and cell cycle of EC109 cells, respectively. RESULTS AND CONCLUSION: With the increased number from 1×104 to 4×104 and increased exposure time from 24 to 72 hours, hEC-MSCs inhibited the proliferation of EC109 cells, and the survival rate of EC109cells was decreased significantly (P < 0.05). Cell cycle analysis revealed a marked increase in the EC-109 G1 phase and a decrease in the EC-109 G2 phase in a number-dependent manner. To conclude, hEC-MSCs inhibit the proliferation of EC109 cells and vary the cell cycle in number- and time-dependent manners.   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 characteristics and differentiation potential of bone marrow embryonic-like stem cells in rats Wang Zheng, Lin Kai 2017, 21 (1):  43-48.  doi: 10.3969/j.issn.2095-4344.2017.01.008 Abstract ( 306 )   PDF (3821KB) ( 270 )   Save BACKGROUND: Rat bone marrow embryonic-like stem cells are found to express embryonic stem cell markers, differentiate into cells derived from three germ layers, and improve cardiac function and remodel the ventricle after implantation. However, the cell biological characteristics and differentiation potential are still unclear. OBJECTIVE: To study the biological properties and differentiation potential of embryonic-like stem cells from rat bone marrow. METHODS: Bone marrow mononuclear cells from 20 Sprague-Dawley were labeled with stage-specific embryonic antigen 1 (SSAE-1), and bone marrow embryonic stem cells were sorted by flow cytometry. Morphology and structure of the sorted cells were observed under scanning electron microscope. Expression of transcription factor (Oct4, Nanog, Sox2) in bone marrow embryonic-like stem cells was detected by immunofluorescence staining. Bone marrow embryonic-like stem cells were cultured and amplified in the feeder layer of fibroblasts to induce myocardium and endothelial cells, and their specific proteins expression was determined.  RESULTS AND CONCLUSION: SSEA-1 labeled rat bone marrow embryonic-like stem cells were gated based on a diameter of 2-6 μm, and these cells were fusiform, polygonal or circular, with a small size. Under the transmission electron microscope, the cell nucleus was enlarged with less cytoplasm containing a small amount of organelles. Multipotential antigen markers could be detected under the immunocytochemical staining of passage 2 rat bone marrow embryonic-like stem cells. With the increase of the number of passages, the cell proliferation ability exhibited a descending trend. One week after myocardial differentiation of rat bone marrow embryonic-like stem cells, myocardial cells were in the parallel arrangement. Two weeks after myocardial induction, the cell morphology changed significantly, and interconnected cells were observed. Immunofluorescence staining showed that cTnT and Cx43 protein were expressed in the myocardium. One week after endothelial differentiation, the endothelial cells were polygonal and spindle-shaped. Two weeks after endothelial differentiation, immunocytochemical staining showed that VWF and CD31 were expressed in the interconnected endothelial cells. To conclude, these findings indicate that rat bone marrow embryonic-like stem cells have similar biological characteristics as bone marrow mesenchymal stem cells and can express stem cell transcription factors. These cells are a likely to differentiate into myocardial and endothelial cells under certain conditions and have a high clinical value.   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 cell transplantation increases bone mineral density of an ovariectomized rat model of osteoporosis Chen Guang-hua, Huang Gui-zhi, Lin Hao, Wu Hao-jun, Chen Hang 2017, 21 (1):  49-53.  doi: 10.3969/j.issn.2095-4344.2017.01.009 Abstract ( 413 )   PDF (4374KB) ( 701 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can be differentiated into osteoblasts, and migrate to lesion sites after transplantation, which have attracted more and more attentions in the research of osteoporosis. OBJECTIVE: To study the effect of BMSCs transplantation on bone mineral density in an ovariectomized rat model of osteoporosis and its possible mechanism of action. METHODS: Using the whole bone marrow culture method, BMSCs were isolated from the femur and tibia of juvenile Sprague-Dawley rats. Thirty adult female rats were randomly divided into three groups: control group, ovariectomized group, and cell transplantation group, with 10 rats in each group. Osteoporosis models were made in ovariectomized and cell transplantation group. Then, PBS solution containing 3.5×109/L BMSCs at a dose of 80 μL/kg was injected via the tail vein into the rats in the cell transplantation group, while the same volume of PBS solution was injected in the other two groups. Two weeks later, dual-energy X-ray absorptiometry was used to measure bone mineral density of the femur, lumbar spine and whole body. Serum levels of calcium, phosphorus, alkaline phosphatase and osteocalcin were determined. Micro-CT was used for morphometric analysis of the tibial microstructure. RESULTS AND CONCLUSION: After ovariectomy, there was a significant reduction in the bone mineral density of the rat femur, lumbar spine and whole body as well as the levels of serum calcium and osteocalcin; however, the phosphorus and alkaline phosphatase levels were increased significantly (P < 0.05 or P < 0.01). These findings indicate the successful preparation of the osteoporosis rat model. Compared with the ovariectomized group, the bone mineral density of the rat femur, lumbar spine or whole body was significantly higher in the cell transplantation group, and significantly increased serum calcium and osteocalcin and significantly decreased phosphorus and alkaline phosphatase levels were also found in the cell transplantation group (P < 0.05). Additionally, the bone volume fraction, number of trabeculae and thickness of trabecular bone were significantly increased in the cell transplantation group compared with the ovariectomized group (P < 0.05). To conclude, our results show that BMSCs can improve the bone mineral density of osteoporosis rats induced by ovariotomy, and thereby promote new bone formation.   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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Interleukin-1beta regulates stem cell gene and chemokine receptor gene expression of myeloma cell lines via pretreatment on bone marrow mesenchymal stem cells Wu Yu-jiao, Fei Xiao-ming, Ye Wei, Tang Yu, Lei Fang, Zhu Yan 2017, 21 (1):  54-59.  doi: 10.3969/j.issn.2095-4344.2017.01.010 Abstract ( 443 )   PDF (1068KB) ( 342 )   Save BACKGROUND: Many studies have showed that mesenchymal stem cells derived from multiple myeloma patients display a wide range of abnormal biological features, including cytokine secretion, immune regulation and osteogenic differentiation. OBJECTIVE: To investigate the expression of stem cell-associated proteins and chemotaxis-related receptors genes in myeloma cells when co-cultured with pulsed or sustained interleukin-1β treated bone marrow mesenchymal stem cells. METHODS: There were three groups in the experiment: untreated normal bone marrow mesenchymal stem cells as control, bone marrow mesenchymal stem cells only treated with interleukin-1β for 1 day as 1-day group or once a day for 7 days as 7-day group. After 7 days of culture, treated or untreated bone marrow mesenchymal stem cells were employed as coating feeder for the direct co-culture with H929 cells for 3 days. At the end of co-culture, H929 myeloma cells were harvested, the expression of NANOG, SOX2 and OCT-4 was measured by western blot, and mRNA levels of CCR1, CCR2, CCR6 and CXCR4 were detected by real-time PCR. RESULTS AND CONCLUSION: The SOX2 protein levels in H929 cells were significantly higher in both 1-day and 7-day groups than that of control group (P < 0.05). In the respective of chemotaxis-related receptors, the mRNA levels of CCR1 and CCR2 in myeloma cells in 7-day group were significantly higher than those in both control and 1-day groups (P < 0.05). Our in vitro results impose inflammatory cytokine interleukin-1β in bone marrow microenvironment has direct effect on myeloma cells, but also regulates myeloma cells indirectly via non-myeloma cells such as mesenchymal stem cells; besides, sustained elevated interleukin-1β level in the bone marrow has more profound effects on myeloma cells through mesenchymal stem cells. Mesenchymal stem cells act an important cellular component involved in myeloma progression.   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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Transforming growth factor beta1 gene silenced adipose-derived stem cell transplantation reduces thrombocytopenic purpura Yin Wan-yi, Shen Yang, Liu Qing-chi, Jia Xiao-hui, Zhang Li-hong 2017, 21 (1):  60-65.  doi: 10.3969/j.issn.2095-4344.2017.01.011 Abstract ( 314 )   PDF (1071KB) ( 512 )   Save BACKGROUND: Some studies have shown that stem cell therapy offers new promise for the treatment of thrombocytopenic purpura, but there are still difficulties in the perfect treatment for thrombocytopenic purpura. OBJECTIVE: To investigate the effect of transplantation of transforming growth factor beta1 (TGF-β1) gene silenced mouse adipose-derived stem cells (ADSCs) in mice with thrombocytopenic purpura. METHODS: The mouse ADSCs were cultured in vitro and transfected by the siRNA expression plasmids containing TGF-β1 (TGF-β1-siRNA). Mouse model of thrombocytopenic purpura were made by guinea pig anti-mouse platelet serum intraperitoneally injected. Then, model mice were randomly assigned into model, ADSCs, TGF-β1 silenced ADSCs, or prednisone group, followed by intravenous injection of 20 μL normal saline, 2×106/L CM-Dil-labeled ADSCs, 20 μL TGF-β1 silenced ADSCs suspension via the tail vein, or intragastrical administration of 5 mg/kg prednisone, respectively. The mice in each group were sacrificed after 14 days of treatment. RESULTS AND CONCLUSION: Compared with the other three groups, the TGF-β1 silenced ADSCs group showed a significant increase in the number of CM-Dil positive cells, blood platelet count, and proportion of thromocytogenic megakaryocytes, while a marked decrease in the total number of megakaryocytes, PAIgG values and TGF-β1 level (P < 0.05). To conclude, TGF-β1 gene silenced ADSCs transplantation can effectively reduce the TGF-β1 and PAIgG values in ITP mice, and promote the differentiation and maturation of megakaryocytes in the bone marrow, which can effectively improve the number of platelets, playing a role in the treatment of ITP.   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 bone marrow mesenchymal stem cell transplantation on proliferation of testicular interstitial cells and secretion of testosterone Liu Ming-kai, Yang Wen-zeng, Ma Tao, Wei Nuo-jing 2017, 21 (1):  66-70.  doi: 10.3969/j.issn.2095-4344.2017.01.012 Abstract ( 422 )   PDF (4160KB) ( 669 )   Save BACKGROUND: Little is known about stem cell transplantation for treatment of partial androgen deficiency syndrome. OBJECTIVE: To observe the effects of bone marrow mesenchymal stem cell transplantation on rat testicular interstitial cells and the secretion of testosterone. METHODS: Thirty Sprague-Dawley rats, 6 months of age, were selected as control group, and serum testosterone and free testosterone concentrations were detected. Another 20 Sprague-Dawley rats, 20 months of age, presenting with lower serum testosterone and free testosterone concentrations than the control group were selected as animal models of partial androgen deficiency syndrome. Then, model rats were randomized into model, orthotopic transplantation, and tail vein transplantation groups. Rats in the latter two groups were respectively given 2×108/L bone marrow mesenchymal stem cell suspension (0.2 mL) into the testis, 2×108/L bone marrow mesenchymal stem cell suspension (0.5 mL) into the tail vein. Eight weeks after transplantation, follicle stimulating hormone, luteinizing hormone and testosterone levels in serum were detected in all groups, and pathological sections were prepared. RESULTS AND CONCLUSION: There was no difference in follicle stimulating hormone and luteinizing hormone levels among the four groups, but significantly lower level of testosterone was found in the model group compared with other groups (P ≤ 0.01). Hematoxylin-eosin staining results showed that: in the normal control group, the testicular tissue was intact, in which, there were a large number of Leydig cells, Sertoli cells and spermatogenic cells; in the model group, the testicular tissue was damaged, with the sparse presence of Sertoli cells and Leydig cells and lack of spermatogonia and spermatogenic cells; in the two transplantation groups, the complete testicular tissue was visible, with a large number of Leydig cells and Sertoli cells or spermatogenic cells. NBT staining results also showed that bone marrow mesenchymal stem cell transplantation increased the testicular interstitial cell number in the testis (P ≤ 0.01). All these findings suggest that bone marrow mesenchymal stem cell transplantation can promote the repair of testicular tissue in old rats, which may lead to a new therapy for partial androgen deficiency syndrome.   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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Adipose mesenchymal stem cells for treatment of traumatic brain injury Zhu Jun-qing, Hong Jun, Cui Jian-zhong, Wang Kai-jie 2017, 21 (1):  71-76.  doi: 10.3969/j.issn.2095-4344.2017.01.013 Abstract ( 314 )   PDF (4493KB) ( 270 )   Save BACKGROUND: Clinical and animal studies have confirmed that transplanted mesenchymal stem cells can migrate to the area of brain injury, and exert a certain therapeutic effect on traumatic brain injury. OBJECTIVE: To study the therapeutic effect of adipose mesenchymal stem cells on the local damage induced by traumatic brain injury in rats. METHODS: According to the literatures, the brain injury model of Sprague-Dawley rats was made by brain freezing injury. After successful modeling, the rats were randomly divided into three groups: model rats in experimental group were administrated with transplantation of adipose mesenchymal stem cells via the tail vein at 2 days after modeling; model rats in control group given the same amount of normal saline; and normal rats in normal group given no treatment. Morris water maze test was used to evaluate the recovery of neurological function in rats. The isolated and cultured adipose mesenchymal stem cells were observed under an inverted microscope. The distribution of these cells in the injured brain and the levels of brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) were detected by immunohistochemistry method. RESULTS AND CONCLUSION: Morris water maze test results showed that compared with the control group, the average escape latency of rats decreased rapidly in the experimental group, and the difference was statistically significant (P < 0.05), while the escape latency of rats in the experimental group was gradually close to that in the control group. Immunohistochemical findings showed that Brdu-labeled adipose mesenchymal stem cells were accumulated and distributed unevenly in the injured cerebral cortex. In the control group, there was no BrdU immunofluorescence staining in the rat brain tissues. Western blot test results showed that: compared with the control group, the BDNF and GDNF levels in the hippocampus of rats were significantly higher in the experimental group (P < 0.05), but compared with the normal group, the BDNF level in the hippocampus of rats was significantly increased, and the GDNF level in the cortex decreased significantly in the control group (P < 0.05). These findings indicate that adipose mesenchymal stem cells can migrate to the damaged area of rats, and promote the secretion of BDNF and GDNF in the injured brain, which may be one of the mechanisms by which adipose mesenchymal stem cell transplantation improves neurological functional recovery of 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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Local effects of adipose-derived mesenchymal stem cells injected into the degenerative intervertebral discs Shen Xiang, Xu Hong-guang, Ma Ming-ming, Zhang Qian, Xiao Liang, Suo Qi-feng 2017, 21 (1):  77-81.  doi: 10.3969/j.issn.2095-4344.2017.01.014 Abstract ( 448 )   PDF (3983KB) ( 255 )   Save BACKGROUND: Domestic studies have shown that adipose-derived mesenchymal stem cells can differentiate into osteoblasts, osteoblasts and other types of bone cells under certain conditions, which are used to repair damaged bone or cartilage tissue. OBJECTIVE: To observe the effect of adipose-derived mesenchymal stem cells on rabbit intervertebral disk degeneration. METHODS: Thirty-six adult New Zealand rabbits were randomized into normal (no treatment), control and experimental groups. Rabbit models of intervertebral disk degeneration were made in the latter two groups using nucleus pulposus aspiration method. Fourteen days after modeling, model rabbits were given injection of BrdU-labeled allogeneic adipose-derived mesenchymal stem cell suspension (50 μL, 1×109/L) in the experimental group or same volume of normal saline in the control group. Thereafter, MRI, hematoxylin-eosin staining and immunohistochemical staining were performed at 2, 4, 8 weeks after injection. RESULTS AND CONCLUSION: MRI findings showed: unchanged disc height and clear and uniform T2-weighted intervertebral disk signals in the normal group; narrowed intervertebral space and weakened T2-weighted signals in the control group that occurred at 4 weeks and gradually developed with time; and increased disc height and T2-weighted signals in the experimental group. Intervertebral disk degeneration shown by hematoxylin-eosin staining occurred at 2 weeks after injection and aggravated in the control group, but in the experimental group, the degeneration was alleviated significantly compared with the control group. Additionally, the transplanted BrdU-labeled adipose-derived mesenchymal stem cells survived and proliferated in the rabbit intervertebral discs. To conclude, adipose-derived mesenchymal stem cells contribute to the recovery from intervertebral disk 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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Umbilical cord mesenchymal stem cells combined with bone marrow stem cells for treatment of lower limb ischemia Du Jun-wen, Wu Tao, Zhang Kun, Su Bai-yu, Lu Cai-ping, Wang Wei-chao, Lei Lin, Guo Jing-xia 2017, 21 (1):  82-86.  doi: 10.3969/j.issn.2095-4344.2017.01.015 Abstract ( 355 )   PDF (4572KB) ( 353 )   Save BACKGROUND: Bone marrow stem cell transplantation has made great progress in the treatment of ischemic disease of the lower limbs. But for the senile patients, the therapeutic outcomes are greatly influenced by few bone marrow stem cells, difficulty in cell collection, and low differentiation ability. OBJECTIVE: To study the clinical efficacy of umbilical cord mesenchymal stem cells combined with bone marrow stem cells in the treatment of lower limb ischemia. METHODS: Forty-seven patients with lower limb ischemia treated with stem cell transplantation in the First Hospital of Shijiazhuang from March 2012 to November 2015 were assigned into either observation group (n=23; transplantation of umbilical cord mesenchymal stem cells combined with bone marrow stem cells) or control group (n=24; bone marrow stem cell transplantation). RESULTS AND CONCLUSION: The total effective rate in the observation group was significantly higher than that of the control group (P=0.039). The skin temperature, percutaneous oxygen branch pressure and ankle brachial index in the observation group were significantly higher than those in the control group (P < 0.05). Scores on limb pain, limb coldness and intermittent claudication in the observation group were significantly lower than those in the control group (P< 0.05). It can be seen that the transplantation of umbilical cord mesenchymal stem cells combined with bone marrow stem cells have better clinical effects on lower limb ischemia stem than single use of bone marrow stem cells, as the combined transplantation can be more effective to relieve clinical symptoms, such as pain and sense of coldness, by improving limb skin temperature, transcutaneous oxygen partial pressure and ankle brachial index.   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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Taurine combined with umbilical cord blood mesenchymal stem cell transplantation for liver injury repair Li Hai-tao, Chen Chao 2017, 21 (1):  87-91.  doi: 10.3969/j.issn.2095-4344.2017.01.016 Abstract ( 384 )   PDF (986KB) ( 326 )   Save BACKGROUND: Studies have shown that umbilical cord mesenchymal stem cells (UC-MSCs) can be induced to differentiate into liver cells, playing a role in liver repair. OBJECTIVE: To explore the effect of taurine combined with UC-MSCs transplantation on liver injury repair. METHODS: Forty healthy Sprague-Dawley rats were used to make chronic alcoholic liver injury models through intragastric administration of 56 degrees liquor, and then randomized into model group, UC-MSCs group and combined treatment group (taurine combined with UC-MSCs transplantation). Model rats were given portal vein injection of 1 mL normal saline in the model group, 1 mL UC-MSCs suspension in the UC-MSCs group, and 0.28 g/L taurine plus 1 mL UC-MSCs in the combined treatment group. Two weeks after transplantation, levels of serum superoxide dismutase, glutathione peroxidase, catalase, alanine aminotransferase, malondialdehyde, endotoxin and interleukin-4 were detected. RT-PCR was used to detect CYP2B1 mRNA expression. RESULTS AND CONCLUSION: Levels of serum superoxide dismutase, glutathione peroxidase, catalase and alanine aminotransferase were highest in the combined treatment group, higher in the UC-MSCs group and lowest in the model group (P < 0.05). The levels of malondialdehyde, endotoxin and interleukin-4 were ranked as follows: combined treatment group < UC-MSCs group < model group (P < 0.05). RT-PCR findings showed that the CYP2B1 mRNA expression was increased significantly in the combined treatment group as compared with the UC-MSCs and model groups (P < 0.05). Taken together, our experimental findings suggest that UC-MSCs transplantation, especially in combination with taurine, can promote recovery from liver injury in rats, which is probably related to a reduction in free radicals that trigger a damage to the body.   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 membrane mesenchymal stem cell transplantation on ovariectomized osteoporosis Lei Ming, Liu Chun-ying, Liu Xiao-feng, Sun Lin, Zheng Wen-kui 2017, 21 (1):  92-97.  doi: 10.3969/j.issn.2095-4344.2017.01.017 Abstract ( 372 )   PDF (1086KB) ( 556 )   Save BACKGROUND: There are rare studies directly exploring the use of human amniotic mesenchymal stem cells to promote differentiation and maturation of osteocytes. Therefore, human amniotic mesenchymal stem cell transplantation is likely to open new horizons for the treatment of osteoporosiss. OBJECTIVE: To explore the effect of human amniotic mesenchymal stem cell transplantation on bone metabolism index and bone microstructure in ovariectomized osteoporosis rats and the relevant mechanisms. METHODS: Thirty-six female Wistar rats without pregnancy were randomly divided into sham and model group. Twenty-three rats in the model group were ovariectomized, and thirteen rats in the sham group received no treatment. Three months after ovariectomy, three rats from each group were executed. ELISA was used to detect serum levels of tartrate-resistant acid phosphatase (TRACP) and bone alkaline phosphatase (BALP). Hematoxylin-eosin staining was performed to observe the pathological changes of the bone. After successful modeling, the remaining rats in the model group were randomized into control and treatment groups, respectively given tail vein injection of sodium chloride and human amniotic mesenchymal stem cells, twice a week, for consecutive 6 weeks. Six weeks after cell transplantation, ELISA was used to detect serum levels of TRACP and BALP; RT-PCR and western blot assay were used to detect expressions of Runx2 and Osterix mRNA and protein; and hematoxylin-eosin staining was used to observe bone microstructure changes. RESULTS AND CONCLUSION: At 3 months after ovariectomy, the rats in the model group exhibited significantly decreased BALP and remarkably increased TRACP as well as sparse number of bone trabeculae some of which were even ruptured shown by hematoxylin-eosin staining. However, the bone trabeculae of rats in the sham group still remained intact. These results indicated the successful modeling in the model group. At 6 weeks after cell transplantation, significantly increased BALP, Runx2, Osterix and significantly decreased TRACP were observed in the treatment group compared with the control group (P < 0.05). Additionally, there a wide-range formation of new bone trabeculae in the treatment group, and damage to the trabecular structure was milder in the treatment group than the control group. In summary, human amniotic mesenchymal stem cell transplantation exerts therapeutic effects on ovariectomized osteoporosis, probably through promotion of Runx2 and Osterix.   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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Combination of olfactory ensheathing cell transplantation and repetitive transcranial magnetic stimulation for the treatment of spinal cord injury Jiang Yong, Chi Xiao-fei, Zou Xi-jun, Ci Yuan, Yao Qi, Yang Mao-wei 2017, 21 (1):  98-102.  doi: 10.3969/j.issn.2095-4344.2017.01.018 Abstract ( 397 )   PDF (2188KB) ( 291 )   Save BACKGROUND: Conventional wisdom suggests that the mammalian brain and spinal cord cannot be regenerated after injury and the hope for functional recovery is slim after spinal cord injury. OBJECTIVE: To test and analyze the therapeutic effect of olfactory ensheathing cell transplantation combined with repetitive transcranial magnetic stimulation (rTMS) in spinal cord injury rats.  METHODS: Eighty rats were used to make spinal cord injury models by T10 laminectomy and then rat models were equivalently randomized into model, cell transplantation, rTMS, and combined treatment groups. Twenty-four hours after modeling, rats in the latter three groups were given olfactory ensheathing cell transplantation around the injured region, rTMS above the left anterior bregma, or their combination, respectively. RESULTS AND CONCLUSION: At 4 weeks following transplantation, the motor function of rats in the combined treatment group was significantly improved, accompanied by increased Basso, Beattie and Bresnahan score and significantly improved sciatic nerve electrophysiological function. These findings indicate that the olfactory ensheathing cell transplantation combined with rTMS treatment achieves remarkable outcomes, and the combined treatment can significantly improve the axonal regeneration and functional recovery after spinal cord 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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Effects of hypoxia inducible factor 1alpha gene modified cardiac stem cell transplantation on cardiac function and survival rate of transplanted cells in rats with myocardial infarction Li Sha, Li Shu-ren, Hao Qing-qing 2017, 21 (1):  103-109.  doi: 10.3969/j.issn.2095-4344.2017.01.019 Abstract ( 317 )   PDF (2371KB) ( 410 )   Save BACKGROUND: The biggest obstacle after stem cell transplantation is the extremely low survival rate of transplanted cells in the severe environments such as ischemia and hypoxia. In order to solve this problem, gene modification is expected to fundamentally improve the survival, metabolism and proliferation/differentiation of stem cells, namely by anti-apoptotic gene transfection or transduction prior to transplantation to vary the gene structure. OBJECTIVE: To observe the survival rate of transplanted cells and cardiac function in rats undergoing hypoxia inducible factor-1α (HIF-1α) gene modified cardiac stem cell transplantation into the edge area of myocardial infarction. METHODS: Twenty-four Sprague-Dawley rats were used to make myocardial infarction models by ligation of coronary arteries and then randomized into HIF-1α-transfected cell transplantation group, cell transplantation group and model group (n=8 per group). Two weeks after modeling, model rats were given the injection of Ad-HIF1α-eGFP-transfected cardiac stem cells, Ad-eGFP-transfected cardiac stem cells or PBS, respectively. RESULTS AND CONCLUSION: Cardiac stem cell transplantation improved the rat cardiac function, and HIF-1α transfected cardiac stem cell transplantation showed better effects on cardiac function improvement than simple cell transplantation. Additionally, the improvement in left ventricular ejection fraction was associated with the density of cardiac stem cells in the marginal zone. To conclude, HIF-1α transfected cardiac stem cell transplantation can effectively improve the cardiac function in rats with myocardial infarction.   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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Feasibility of renal tissue stem cell therapy for acute renal failure Hu Yue-shi, Li Peng, Cao Zhi-hua, Liu Lei, Wang Yang 2017, 21 (1):  110-114.  doi: 10.3969/j.issn.2095-4344.2017.01.020 Abstract ( 512 )   PDF (3403KB) ( 451 )   Save BACKGROUND: Studies have shown that renal tissue stem cells mainly exist in the renal papillae, holding similar characteristics to mesenchymal stem cells, yet it is unknown whether they are involved in the renal repair of tubular epithelial injury. OBJECTIVE: To explore the effect and feasibility of renal tissue stem cells in the repair of acute renal failure. METHODS: Renal tissue stem cells were isolated and culture from the renal tissue of 10 Sprague-Dawley rats. Another 30 Sprague-Dawley rats were randomly assigned into experimental group, model group or normal control group. An animal model of acute renal failure was made in the experimental and model groups. Six hours after modeling, experimental rats were respectively given tail vein injection of renal tissue stem cells in the experimental group, given the same volume of normal saline in the model group, and given no treatment in the normal control group. Ten days after transplantation, the renal function was detected, and renal tissue samples were taken and observed histologically. RESULTS AND CONCLUSION: After transplantation, levels of urea nitrogen and creatinine were improved significantly in the experimental group (P < 0.05), which were similar to those in the normal control group (P > 0.05). No abnormal changes and no inflammatory infiltration in the cortex and outer medullary tubules were found in the normal control group, while in the experimental group, inflammatory cell infiltration and epithelial cell swelling were visible but milder than the model group. To conclude, our experimental findings show that the renal tissue stem cells get ideal effects on the renal function of acute renal failure rats by promoting epithelial cell 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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Effects of recombinant adenovirus-mediated adiponectin on human umbilical vein endothelial cell injury and the underlying mechanism Wang Xue-mei, Wei Qin, Duan Ming-jun, Zhang Chun, Yang Yi-ning 2017, 21 (1):  115-121.  doi: 10.3969/j.issn.2095-4344.2017.01.021 Abstract ( 347 )   PDF (4992KB) ( 661 )   Save BACKGROUND: Vascular endothelial cells closely adhere to the blood vessel wall so as to play a crucial role in the regulation of vascular homeostasis. OBJECTIVE: To analyze the inhibitory effects of recombinant adenovirus-mediated adiponectin overexpression on oxidized low density lipoprotein (ox-LDL)-induced atherosclerotic injury in human umbilical vein endothelial cells, nuclear factor κB (NF-κB) p65 nuclear protein and pro-inflammatory factors in the downstream of signaling pathway. METHODS: There were four groups in this experiment: human umbilical vein endothelial cells were cultured in the conventional medium without ox-LDL induction as control group; human umbilical vein endothelial cells induced by 30 mg/L ox-LDL for 48 hours as ox-LDL induction group; cells firstly treated with 100 µmol/L pyrrolidine dithiocarbamate (PDTC) for 1 hour, and then induced by 30 mg/L ox-LDL for 48 hours as PDTC with ox-LDL induction group; cells transfected by adenovirus bearing a vector encoding for adiponectin and enhanced green fluorescent protein (Ad-APN-eGFP) (multiplicity of infection=100 pfu/cell) for 2 hours, and then induced by 30 mg/L ox-LDL for 48 hours as Ad-APN-eGFP with ox-LDL induction group. Afterwards, all relative indicators were detected in each group. RESULTS AND CONCLUSION: Endothelial cell injury models were obtained by ox-LDL induction, and the NF-κB signaling pathway was activated with the increased expression of NF-κB p65 nuclear protein. Ad-APN-eGFP could inhibit the NF-κB signaling pathway, which significantly decreased the expression of NF-κB p65 nuclear protein, as well as the gene and protein expressions of VCAM-1, ICAM-1 and MCP-1 in the downstream of signaling pathway, but significantly increased the gene and protein expressions of endothelial nitric oxide synthase. These results show that Ad-APN-eGFP-tranfected human umbilical vein endothelial cells can relieve ox-LDL-induced pro-inflammatory injury and reduced apoptosis via effectively inhibiting the NF-κB activation, which achieves similar outcomes with the PDTC intervention. In other words, Ad-APN-eGFP and PDTC both can reduce inflammatory reactions such as atherosclerotic injury by inhibiting the NF-κB signaling 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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Directional differentiation of human amniotic mesenchymal stem cells into osteoblasts, chondrocytes and lipocytes in vitro Li Yu-wan, Zhu Xi-zhong, Jin Ying, Wang Sheng-min, Yang Ji-bin, Xiong Hua-zhang, Zou Gang, Liu Yi 2017, 21 (1):  122-127.  doi: 10.3969/j.issn.2095-4344.2017.01.022 Abstract ( 398 )   PDF (5026KB) ( 335 )   Save BACKGROUND: Human amniotic mesenchymal stem cells (hAMSCs) are a kind of adult stem cells that can differentiate into osteocytes, chondrocytes and lipocytes under different conditions in vitro. Compared with other mesenchymal stem cells (MSCs), hAMSCs have many advantages like wide source, noninvasive isolation, low immunogenicity and short growth cycle, making hAMSCs an important source of seed cells in tissue engineering. OBJECTIVE: To investigate whether hAMSCs have the characteristics of MSCs and the ability of differentiation  into osteoblasts, chondrocytes and lipocytes. METHODS: Agreed by patients, the amniotic membrane from partus maturus parturients was separated, and hAMSCs were isolated by enzyme digestion. Passage 3 cells were cultured in osteogenic, chondrogenic or adipogenic medium induction. RESULTS AND CONCLUSION: The alizarin red staining results showed multiple mineralized nodules after osteogenic induction, and the activity of alkaline phosphatase was significantly increased after induction. The toluidine blue staining showed positive results after chondrogenic induction, and a large amount of glycosaminoglycans were secreted from the cell matrix. After adipogenic differentiation, lipid droplets were observed in the cytoplasm of the cells shown by the oil red O staining. Real-time PCR results showed the mRNA expressions of relative genes at day 21 after osteogenic, chondrogenic and adipogenic induction upregulated significantly compared with the results at day 7 (P < 0.05). These experimental results demonstrate that hAMSCs have the basic characteristics of MSCs and possess the potential of differentiation into osteoblasts, chondrocytes and lipocytes, which could be selected as the seed cells for 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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Effects of exogenous stem cell factors on the gastric electrical activity and impedance in a mouse model of diabetic gastroparesis Gao Jin-liang, Zhao Nan, Song Zhan 2017, 21 (1):  128-132.  doi: 10.3969/j.issn.2095-4344.2017.01.023 Abstract ( 347 )   PDF (3768KB) ( 454 )   Save BACKGROUND: Lesions in Cajal interstitial cells can lead to diabetic gastroparesis, in which, insulin and stem cell factor are likely to act as upstream regulatory factors. OBJECTIVE: To investigate the effect of exogenous stem cell factors on the gastric electrical activity and gastric impedance in a mouse model of diabetic gastroparesis. METHODS: Forty Wistar rats were randomly divided into two groups, 20 rats in each group. The diabetic gastroparesis model was prepared in observation group by a single use of 60 mg/kg streptozotocin via intraperitoneal injection and high-sugar/high-fat diet. Rats in control group were given the equal volume of normal  saline and normal diet. Eight weeks later, residual rate of gastric pigment was detected, and gastric antrum microstructures were observed under transmission electron microscope. If the animal model was successfully prepared, rats were intraperitoneally given 0.4 μg/kg/d stem cell factor in the observation group and same volume of PBS in the control group, for consecutive 15 days. Afterwards, the impedance gastric motility system was detected. RESULTS AND CONCLUSION: Compared with the control group, the residual rate of gastric pigment was significantly higher in the observation group (P < 0.05). In the observation group, there was a certain change in the Cajal interstitial cells in the antrum of stomach, such as the gap junction between the cells were significantly reduced and became loose; there was a lack of continuity of the basal lamina; the number of organelles in the cells was significantly decreased, the cytoplasm was dissolved, and a large number of vacuoles appeared. After treatment with exogenous stem cell factor, the gastric electrical activity and gastric impedance of rats were significantly improved: for the gastric electrical activity, the percentage of < 2 band power was significantly decreased, and the percentage of 2-4 band power increased significantly (P < 0.05); for the gastric impedance, the percentage of < 2 band power was significantly decreased, and the percentage of 2-4 band power increased significantly (P < 0.05). After treatment, no significant difference in the gastric electrical activity and gastric impedance was found between the two groups. The experimental results show that delayed gastric emptying may occur in diabetic gastroparesis rats, and the degeneration of Cajal interstitial cells may induce gastroparesis. Exogenous stem cells can effectively improve the level of gastric electrical activity and gastric impedance, and thereby improve the gastric motility 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 corbrin capsule combined with human amniotic mesenchymal stem cell transplantation on rat renal function and hypercoagulability Guo Zhi-bo, Zhang Chen-jie, Ma Li-na, Gao Da-wei, Zhou Fang, Li Na-na, Zhou Jun-xue, Zhang Na 2017, 21 (1):  133-139.  doi: 10.3969/j.issn.2095-4344.2017.01.024 Abstract ( 320 )   PDF (1257KB) ( 260 )   Save BACKGROUND: As the main component of cordycepic acid, corbrin capsule mainly functions to improve blood lipids and protect the kidneys. Human amniotic mesenchymal stem cells are a kind of stem cells that have stronger differentiation potential than bone marrow mesenchymal stem cells. However, there are less reports on renal injury repair and post-repair functional improvement. OBJECTIVE: To observe the effect of human amniotic mesenchymal stem cell transplantation with corbrin capsule on blood coagulation and renal functions of rats with nephrotic syndrome. METHODS: Human amniotic mesenchymal stem cells were cultured in vitro and made into cell suspension labeled with with CM-Dil. Fifty Sprague-Dawley were randomly divided into five groups (n=10 per group): normal control group with no intervention, model group, cell transplantation group, corbrin capsule group and combined treatment group. Except the normal control groups, rats in all the groups were used to make nephropathy models by rats by intravenous injection of adriamycin. After modeling, rats in the latter three groups were given intravenous injection of human amniotic mesenchymal stem cells for 3 days, intragastric administration of corbrin capsule for 12 weeks, and the combination of cell transplantation and corbrin capsule, respectively. Twelve weeks later, serum creatinine, blood urea nitrogen, blood coagulation index, and 24-hour urine protein levels were detected. Pathological changes of the renal tissues were observed by hematoxylin-eosin staining. Survival and distribution of CM-Dil labeled human amniotic mesenchymal stem cells were observed by fluorescence microscope. Apoptosis in renal tissues was detected using TUNEL method. RESULTS AND CONCLUSION: Serum creatinine, blood urea nitrogen and 24-hour urine protein levels in the cell transplantation and corbrin capsule groups were significantly decreased compared with the model group, but still higher than that in the combined treatment group (P < 0.05). Coagulation indexes in the combined treatment group were significantly lower than those in the cell transplantation and corbrin capsule groups (P < 0.05). The hypercoagulable state of rats was relieved in the three treatment groups. Pathological changes of the renal tissues were deteriorated in the model group, and improved in the three treatment groups, especially in the combined treatment group. Under the florescence microscope, the number of CM-Dil-labeled human amniotic mesenchymal stem cells was higher in the combined treatment group than the cell transplantation and corbrin capsule groups. Compared with the model group, the cell apoptosis in the renal tissue was relieved in the three treatment groups, especially in the combined treatment (P < 0.05). To conclude, human amniotic mesenchymal stem cell transplantation with corbrin capsule can improve renal function and hypercoagulable state of nephrotic syndrome rats, by reducing the degree of proteinuria, improving the degree of pathological damage and reducing renal cell apoptosis.   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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Combined effects of neural stem cell transplantation and electroacupuncture on the behaviors of chronic stress-induced depression rats Mi Kun, Guo Qiang, Sun Zhen-qing, Wang Hai-bo 2017, 21 (1):  140-145.  doi: 10.3969/j.issn.2095-4344.2017.01.025 Abstract ( 258 )   PDF (1092KB) ( 229 )   Save BACKGROUND: Both electroacupuncture and neural stem cell transplantation have certain therapeutic effects on chronic stress depression, but their synergistic effects remain unclear. OBJECTIVE: To explore the effect of electroacupuncture intervention combined with neural stem cell transplantation via tail vein on the behaviors of chronic stress-induced depression rats. METHODS: Fifteen of 63 rats were randomly selected as normal group, and the other 48 rats were fed individually and underwent chronic unpredictable mild stress stimulation to make rat models of depression. Except for three unsuccessful models, 45 successful models were randomly assigned into model (no intervention), neural stem cell (injection of 1 mL neural stem cell suspension (3×106) via the tail vein), or combined treatment group (injection of 1 mL neural stem cell suspension (3×106) via the tail vein+1-hour electroacupuncture every other day for consecutive 1 week) (n=15/group). The behavior ability of rats was assessed through sucrose preference test and open-field test. The survival, distribution and apoptosis of neural stem cells were assessed by CM-Dil labeling and TUNEL assay, respectively. The expression of Caspase-3 and Bcl-2 in the hippocampal CA3 region was assessed by RT-PCR and western blot assay. RESULTS AND CONCLUSION: Compared with the normal group, model group rats exhibited lower body mass, lower scores on the sucrose preference test and open-field test, higher Caspase-3 mRNA and protein expressions (P < 0.05), lower Bcl-2 protein and mRNA expressions (P < 0.05) and more apoptotic neurons (P < 0.05). Compared with the model group, rats in the neural stem cell group and combined treatment group had significant increases in the body mass and scores on the sucrose preference test and open-field test as well as exhibited significantly decreased Caspase-3 mRNA and protein expressions (P < 0.05) and increased Bcl-2 protein and mRNA expressions (P < 0.05). These indicators were improved better in the combined treatment group than the neural stem cell group (P < 0.05). Under the fluorescence microscope, there were more CM-Dil-positive cells in the combined treatment group than the neural stem cell group. To conclude, neural stem cell transplantation can improve depressive behaviors in rats with chronic stress-induced depression, and electroacupuncture can elevate the survival rate of transplanted neural stem cells by reducing Caspase-3 and increasing Bcl-2 expression.   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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Dental pulp stem cells in the paradigm of migration Sidra Fahim, Pan Shuang, Zhang Lin, Niu Yu-mei 2017, 21 (1):  146-153.  doi: 10.3969/j.issn.2095-4344.2017.01.026 Abstract ( 316 )   PDF (1155KB) ( 583 )   Save BACKGROUND: Continuing characterization of a multitude of signaling pathways associated with migration suggests that modification of migration in dental pulp stem cells can be achieved in a way which suits efficient tissue regeneration. OBJECTIVE: To identify and analyze the published work undertaken regarding different aspects of migration of dental pulp stem cells. METHODS: A PubMed, ScienceDirect (Elsevier), EBSCOhost, Wiley Online Library, MEDLINE, Scholar Universe, PMC and OVID database search was conducted to access the publications spanning over a period of 15 years ranging from the year 2000 to 2015 by two independent reviewers. The keywords used in our search were “dental pulp”, “stem cells”, “migration”, “pulpal injury”. Language preference was set for articles in English language in this research. This search strategy yielded a total of 46 citations from which relevant studies were selected for review according to pre-determined inclusion/exclusion criteria. RESULTS AND CONCLUSION: For the final review process, 38 articles pointing towards major findings determining migratory behavior of dental pulp stem cells under the effect of a variety of biochemical stimuli and dental materials used in clinical work were retrieved. This review article highlights an implication upon usage of dental pulp stem cells for tissue engineering practices.   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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Molecular mechanism and signaling pathway during adipogenic differentiation of adipose-derived stem cells Chen You-bai, Hao Yong-hong, Wang Lan, Zhang Qi-xu, Han Yan 2017, 21 (1):  154-158.  doi: 10.3969/j.issn.2095-4344.2017.01.027 Abstract ( 778 )   PDF (976KB) ( 373 )   Save BACKGROUND: Research on adipogenic differentiation of adipose-derived stem cells (ADSCs) cannot only help us to explore the process and mechanism of obesity, but also to construct tissue-engineered adipose tissue which providing a new method for soft tissue reconstruction. OBJECTIVE: By summarizing the process, molecular mechanism and signaling pathway, to conclude the effect of miRNA on ADSCs adipogenic differentiation, and to discuss the relation between adipogenic and osteogenic differentiations of ADSCs. METHODS: Relevant articles were searched in PubMed and SinoMed using the following keywords of “adipose stem cell, adipose-derived stem cell, adipose-derived mesenchymal stem cell, differentiation” in English and Chinese, respectively. Finally, 27 representative articles were included. RESULTS AND CONCLUSION: PPARγ/MAPK, PI3K/Akt, Wnt/β-Catenin, cAMP, and Notch signal pathways play important roles in adipogenic differentiation of ADSCs; however the relevant genes, proteins, signaling pathways and underlying molecular mechanisms are not well understood. miRNAs have great influence on adipogenic differentiation of ADSCs. For example, miR-24, miR-148a and miR-302 can promote and miR-22, miR-27 can inhibit the adipogenic differentiation of ADSCs. The osteogenic and adipogenic differentiations of ADSCs are closely related. Some growth factors and hormone facilitate the adipogenic but inhibit the osteogenic differentiation of ADSCs simultaneously, whereas others have opposite effects. The future investigation may concentrate on comprehensively utilizing various growth factors and scaffolds to mimic in vivo microenvironment and to improve proliferation and adipogenic differentiation of ADSCs for construction of tissue-engineered adipose tissue.   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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Instructions for Authors (2017)-Chinese Journal of Tissue Engineering Research (CJTER) CHINESE JOURNAL OF ENGINEERING RESEARCH 2017, 21 (1):  159-164.  doi: 10.3969/j.issn.2095-4344.2017.01.028 Abstract ( 586 )   PDF (800KB) ( 464 )   Save Related Articles | Metrics