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07 October 2016, Volume 20 Issue 41 Previous Issue    Next Issue

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Effect of fructose and dithiothreitol on cell viability and pluripotency of cryopreserved bone marrow mesenchymal stem cells Zheng Xin-tong, Liu Qin, Zhang Jing-xia, Luo Qing, Chen Zhe, Song Guan-bin 2016, 20 (41):  6085-6091.  doi: 10.3969/j.issn.2095-4344.2016.41.001 Abstract ( 262 )   PDF (1227KB) ( 251 )   Save BACKGROUND: Cell cryopreservation is required for clinical use of stem cells, and the current process of cryopreservation however may be harmful to cell viability, pluripotency and differentiation capacity. OBJECTIVE: To explore the effect of fructose and dithiothreitol on pluripotency and osteogenesis of cryopreserved bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were isolated from the bone marrow of Sprague-Dawley rats and pretreated with fructose (200 μmol/L), dithiothreitol (500 μmol/L) or combined components before cryopreservation. Then the cells were cryopreseved for 6 months and the morphology of cells was observed by inverted microscopy. The cell viability was evaluated by MTT, and real-time PCR was used to detect the mRNA expression of Nanog, OCT4 and Sox2. Alkaline phophatase activity assay and alizarin red staining were utilized to detect the osteogenic capacity of bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: Images captured by inverted microscopy showed no significant difference in cell morphology between groups. The MTT results indicated that fructose and combined pretreatment could promote the cell viability of bone marrow mesenchymal stem cells after cryopreservation, while the real-time PCR results demonstrated that dithiothreitol significantly facilitated the expression of Naogo and Sox2 in bone marrow mesenchymal stem cells. Moreover, ALP activity assay and alizarin red staining confirmed the positive effects of fructose, dithiothreitol and combined pretreatment on osteogenic capacity of bone marrow mesenchymal stem cells after cryopreservation, and the best effects were found after pretreatment with dithiothreitol and combined components. Overall, these findings indicate that fructose pretreatment is beneficial for cell viability of cryopreseved bone marrow mesenchymal stem cells, and dithiothreitol contributes to maintaining the pluripotency and osteogenesis capacity of cryopreseved bone marrow mesenchymal 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 cell passage on differentiation of bone marrow mesenchymal stem cells into neural stem cells Liang Wei, Liu Zhou, Xu Zhi-en, Lin Li-feng, Fang Hong-ming 2016, 20 (41):  6092-6097.  doi: 10.3969/j.issn.2095-4344.2016.41.002 Abstract ( 224 )   PDF (1297KB) ( 234 )   Save BACKGROUND: It is unclear whether serial cell passage in vitro influences the differentiation of bone marrow mesenchymal stem cells into neural stem cells. OBJECTIVE: To investigate the effect of cell passage on the differentiation of bone marrow mesenchymal stem cells into neural stem cells. METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured by the whole bone marrow adherence method. Bone marrow mesenchymal stem cells at passages 3, 6, 9, 12 were incubated in serum-free medium. After culture for 7 and 14 days, cell biological characterization was observed and differenitaiton ability into neural stem cells was observed by detecting Nestin expression in cells using flow cytometry. Then, the cells were further induced to differentiate and cell multipotential differentiation capacity was detected by measurement of nerve enolase and glial acidic protein expression. RESULTS AND CONCLUSION: Under induction, bone marrow mesenchymal stem cells at different passages were all differentiated into Nestin-positive neural stem cells. However, there was a significant difference in differentiation proportion of cells at different passages (P < 0.05). Strongest differentiation ability was found in the passage 6 cells, with the Nestin expression up to (93.7±2.3)% at 7 days of induction and (96.2±1.8)% at 14 days of induction. The proportion of differentiated cells at passages 6 and 9 was signfiicantly higher than that at passages 3 and 12. Moreover, adherent cells were positive for nerve enolase and glial acidic protein. All these findings indicate that the differentiation of bone marrow mesenchymal stem cells into neural stem cells is correlated with cell passage. Cells at lower or higher passages are both detrimental to cell differentiation.     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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Stromal cell derived factor-1/chemokine receptor 4 signaling pathway is involved in bone morphogenetic protein 2-induced migration of bone marrow mesenchymal stem cells Yang Yi, Yi Lei, Yeerzhati Hajiaheman, Telieke Kanzhale, Jin Ge-le 2016, 20 (41):  6098-6104.  doi: 10.3969/j.issn.2095-4344.2016.41.003 Abstract ( 230 )   PDF (5268KB) ( 228 )   Save BACKGROUND: Stromal cell derived factor-1 (SDF-1)/chemokine receptor 4 (CXCR-4) biological axis plays a chemotactic role in a variety of cells, making it possible to regulate the regeneration of a variety of tissues. Whether the bidogical is involved in bone morphogenetic protein-2 (BMP-2)-induced homing of bone marrow mesenchymal stem cells, however, is still unclear. OBJECTIVE: To study the role of SDF-1/CXCR4 signaling pathway in BMP-2-induced migration of mouse bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells in logarithmic growth were selected and intervened with SDF-1 (0, 50, 100 and 200 μg/L), BMP-2 (0, 50, 100 and 200 μg/L) and AMD3100 (50 μg/L) to induce cell migration detected by Transwell method. RESULTS AND CONCLUSION: The migration of bone marrow mesenchymal stem cells was closely related to SDF-1 and BMP-2, and proportional to the concentration of both SDF-1 and BMP-2. When SDF-1 and BMP-2 were used jointly, the number of migrated cells was increased significantly, and highest number of migrated cells was obtained at 200 μg/L. Moreover, these migrated cells showed a nest-like distribution under microscopy. AMD3100 as an inhibitor markedly suppressed the migration of bone marrow mesencnymal stem cells induced by BMP-2, but the number of migrated cells was likely to increase with the increasing concentration of BMP-2 that exceeded a specific value. Overall, our findings show that SDF-1/CXCR4 signaling pathway is an important pathway in BMP-2-induced migration of bone marrow mesenchymal 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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Role of adipose-derived stem cells in the fat transplantation Zhao Xue-lian, Zhang Chun-li, Su Xiao-guang, Zhang Zhuo-nan, Han Peng, Zhang Jie, Wang Yan-ling, Zhang Chui 2016, 20 (41):  6105-6111.  doi: 10.3969/j.issn.2095-4344.2016.41.004 Abstract ( 312 )   PDF (5178KB) ( 310 )   Save BACKGROUND: There are a lot of adipose-derived stem cells in the vascular stroma. These cells are shown to play a very important role in the fat granule transplantation. OBJECTIVE: To explore the role of adipose-derived stem cells in the fat granule transplantation. METHODS: Normal adipose tissues were obtained from 10 male BALB/C mice, SPF grade. Adipose-derived stem cells and fat granules were extracted from the abdominal fat tissues. Another 24 nude mice acted as recipients and were assigned into control, fat granule transplantation or mixed transplantation (adipose-derived stem cells+fat granules) groups. In the latter two groups, fat granule suspension and suspension of fat granules and adipose-derived stem cells were injected into the shoulder of rats, respectively. In the control group, the same volume of cell medium was injected. Four weeks later, separated plasma and grafts were taken out for indicator measurement. RESULTS AND CONCLUSION: Compared with the fat granule transplantation group, the mixed transplantation could remarkably increase the weight of grafts, while reduce the absorption of grafted fat tissues (P < 0.01). After transplantation, the highest level of vascular endothelial growth factor in the plasma was obtained in the mixed transplantation group followed by fat granule transplantation group and control group (P < 0.01). Level of basic fibroblast growth factor and microvessel density were significantly higher in the mixed transplantation group than the fat granule transplantation group (P < 0.01). Better cell morphology and higher number of fat droplets were found in the mixed transplantation group compared with the fat granule transplantation group. All these results indicate that adipose-derived stem cell transplantation can remarkably promote the expression of basic fibroblast growth factor, improve graft microcirculation, and improve morphology and function of fat granules.     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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Correlation between growth, proliferation and apoptosis of leukemia cell lines K562/AO2 and bone marrow mesenchymal stem cells in children with leukemia Lei Chun-xia 2016, 20 (41):  6112-6117.  doi: 10.3969/j.issn.2095-4344.2016.41.005 Abstract ( 268 )   PDF (1048KB) ( 254 )   Save BACKGROUND: Leukemia comes from the damage to bone marrow microenvironment, and changes in bone marrow microenvironment lead to some effects on biological characteristics and behavior of leukemia cells. OBJECTIVE: To explore the effect of bone marrow mesenchymal stem cells from children with leukemia on K562/AO2 cell line growth, proliferation and apoptosis. METHODS: K562/AO2 cells were cultured alone in vitro, or co-cultured with bone marrow mesenchymal stem cells from leukemia children. The living cells in the two groups were counted using trypan blue staining, and the cell growth curves were described. PI single staining method was used to detect the cell cycle, and cell apoptosis was detected by V/PI Annexin method in the two groups. RESULTS AND CONCLUSION: After single suspension culture, the cells showed no marked changes within the first 3 days, but began to proliferate fast at 4 days and reached the proliferation peak at 6 days. Co-cultured cells showed a gentle growth curve with no presence of obvious proliferation peak. Compared with the co-culture group, the proportion of G0/G1 phase cells was significantly lower, but the proportion of S phase cells was significantly higher in the single culture group (P < 0.05). The number of cells in the G2/M phase showed no significant difference between the two groups (P > 0.05). All these findings indicate that co-culture with bone marrow mesenchymal stem cells from leukemia children inhibits the proliferation of K562/AO2 cells through G0/G1 phase arrest, and also resists apoptosis in K562/AO2 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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Paclitaxel and cisplatin inhibit the proliferation of nasopharyngeal cancer stem cells and promote apoptosis via the Wnt/beta-catenin pathway Liu Yong-gang, Yang Rong-song, Wu Hong-fang, Zhang Bao-chao 2016, 20 (41):  6118-6124.  doi: 10.3969/j.issn.2095-4344.2016.41.006 Abstract ( 229 )   PDF (1153KB) ( 242 )   Save BACKGROUND: Cancer stem cells have self-renewal ability and can differentiate into new tumors. Cancer stem cells are the source of tumor formation and recurrence, and they can make tumors insensitive to radiotherapy and chemotherapy. OBJECTIVE: To explore the effect of paclitaxel plus cisplatin on the proliferation and apoptosis of nasopharyngeal cancer stem cells (NPCSCs) and involved signal pathways. METHODS: NPCSCs were sorted by immunomagneticbeads and were treated with paclitaxel, cisplatin or their combination. The expression of caspase-3, activated caspase-3 and Bcl-2, which are related to apoptosis, was determined by western blot. The expression of β-catenin and its downstream proto-oncogene, c-myc, was also determined by western blot. The activity of the Wnt/β-catenin pathway was inhibited by knocking down β-catenin expression or β-catenin inhibitor XAV939. Proliferation and apoptosis of NPCSCs were detected by MTT and flow cytometry, respectively. RESULTS AND CONCLUSION: Either paclitaxel or cisplatin could inhibit proliferation and induce apoptosis of NPCSCs. The expression of apoptosis marker, activated caspase-3, was increased and the expression of the inhibitor of apoptosis, Bcl-2, was declined. Combined use of paclitaxel and cisplatin had synergistic effect when used together. Either paclitaxel or cisplatin could inhibit the expression of β-catenin and c-myc, suppressed the proliferation and induced the apoptosis of NPCSCs by inhibiting the activity of Wnt/β-catenin pathway. These results indicate that the combined use of paclitaxel and cisplatin may inhibit the proliferation of NPCSCs and promote apoptosis via the Wnt/β-catenin 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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Safety assessment of mammary gland stem cells from normal tissues in breast cancer patients Bi Xiao-juan, Fu Ming-gang, Guo Li-ying, Liu Sha, Dilimina Yilamu, Guo Chen-ming 2016, 20 (41):  6125-6130.  doi: 10.3969/j.issn.2095-4344.2016.41.007 Abstract ( 310 )   PDF (4834KB) ( 297 )   Save BACKGROUND: The cultivation of mammary gland stem cells is of great significance for the development of mammary gland and breast cancer. OBJECTIVE: To seek an easy method to isolate and culture mammary gland stem cell in vitro, and verify the safety of cells. METHODS: Mammary epithelial cells were isolated from normal tissues surrounding breast cancer, and CD49f- and EPCAM-positive cells were sorted using flow cytometry followed by surface marker analysis and cell colony formation ability analysis. Afterwards, real-time fluorescent quantitative PCR was used to detect C-erbB-2 and Maspin mRNA expression in mammary gland stem cells, breast cancer tissues and normal tissues surrounding breast cancer. RESULTS AND CONCLUSION: Human mammary gland stem cells were successfully cultured and highly expressed CD49f and EPCAM, with the presence of mixed colony, pleural epithelial cell colony, and myoepithelial cell colony. c-erbB-2 was lowly expressed while Maspin highly expressed in mammary gland stem cells. Our experimental findings indicate that the mammary gland stem cells derived from normal tissue surrounding breast cancer have biological 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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Effect of Acorus tatarinowii on the osteogenic differentiation of umbilical cord blood stem cells Chu Jing, Wang Li-qin, Zhang Han, Zeng Juan 2016, 20 (41):  6131-6137.  doi: 10.3969/j.issn.2095-4344.2016.41.008 Abstract ( 283 )   PDF (4901KB) ( 178 )   Save BACKGROUND: Studies have demonstrated that Acorus tatarinowii and its active ingredients can promote adult neurogenesis, exerting an active role in anti-aging and neurodegenerative disease treatment. OBJECTIVE: To explore the effects of Acorus tatarinowii extracts on the proliferation and osteogenic differentiation of umbilical cord blood stem cells, thereby providing a new idea for promoting the osteogenic differentiation of stem cells by Chinese medicines. METHODS: Acorus tatarinowii extracts were obtained via solvent extraction method and flow cytometry sorting technology was used to select the stem cells isolated from human umbilical cord. Then, the umbilical cord blood stem cell proliferation was observed by electron microscope, and the effect of Acorus tatarinowii on the proliferation of umbilical cord blood stem cells was observed by cell counting kit-8. Meanwhile, the impact of Acorus tatarinowii on osteocalcin and bone morphogenetic protein-2 contents in the supernatant of umbilical cord blood stem cells were detected by ELISA; alkaline phosphatas expression was detected using alkaline phosphatase staining kit. RESULTS AND CONCLUSION: The separation purity of the stem cells from umbilical cord mononuclear cells was (89.66±3.47)%. After low, moderate and high concentrations of cord blood stem cells co-cultured with Acorus tatarinowii extractions for 24, 48 and 72 hours, the stem cell proliferation rate was significantly higher compared with the control group (P < 0.05), and additionally, the proliferation rate of moderate concentration group was significantly higher than that in the low and high concentration groups (P < 0.05). The contents of osteocalcin and bone morphogenetic protein-2 in the stem cell supernatants were significantly higher than those in the control group after co-cultured with Acorus tatarinowii extractions for 5, 10 and 15 days, and which the highest in the high concentration group (P < 0.05). The expression of alkaline phosphatase was significantly higher than that in the control group after umbilical cord blood stem cells co-cultured with Acorus tatarinowii extractions for 10 days, and moreover, the expression of alkaline phosphatase in the moderate concentration and high concentration groups were significantly higher than that in the low concentration group (P < 0.05). In conclusion, Acorus tatarinowii can promote the proliferation and osteogenic differentiation of umbilical cord blood 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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Treatment of traumatic brain injury by Panax notoginseng saponins combined with bone marrow mesenchymal stem cell transplantation Li Lei-bing, Wang Jiao-yue, Sun Cai-hong 2016, 20 (41):  6138-6144.  doi: 10.3969/j.issn.2095-4344.2016.41.009 Abstract ( 230 )   PDF (2237KB) ( 227 )   Save BACKGROUND: Studies have shown that Panax notoginseng saponins (PNS) has extensive pharmacological basis for the treatment of cerebral ischemic injury, and animal experiments for treatment of brain injury by bone marrow mesenchymal stem cell transplantation are ongoing. However, little is reported about the combined use of PNS and bone marrow mesenchymal stem cell transplantation. OBJECTIVE: To investigate the effect of PNS combined with bone marrow mesenchymal stem cell transplantation on traumatic brain injury in rats. METHODS: Traumatic brain injury models were made in 60 Sprague-Dawley rats by hydraulic shock method. Then, model rats were randomized into model group, cell transplantation group receiving bone marrow mesenchymal stem cell transplantation and combined treatment group undergoing PNS combined with bone marrow mesenchymal stem cell transplantation. Nerve function recovery of the rats and protein expression of nerve growth factor in the brain tissue were evaluated and detected by Bederson scoring and western blot methods, respectively. Morphological changes of the brain tissue and apoptosis in cortical neurons were observed and detected by hematoxylin-eosin staining and TUNEL, respectively. RESULTS AND CONCLUSION: Highest Bederson score was found in the model group, followed by the cell transplantation group and combined treatment group (P < 0.05), while the protein expression of nerve growth factor was ranked as follows: the combined treatment group > the cell transplantation group > the model group (P < 0.05). Inflammatory infiltration and brain edema in the brain were relieved markedly in the combined treatment group compared with the other two groups, and the number of apoptotic neurons was significantly reduced as well. Our findings suggest that PNS combined with bone marrow mesenchymal stem cell transplantation can promote neurological recovery from traumatic brain injury by increasing the expression of nerve growth factor and reducing neuronal 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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Interleukin 8 is involved in the invasion and metastasis of CD133+ hepatocellular carcinoma stem cells Wen Li-hong, Hu Wen-jie, Ye Heng-xi, Xiang Wei-dong 2016, 20 (41):  6145-6150.  doi: 10.3969/j.issn.2095-4344.2016.41.010 Abstract ( 257 )   PDF (1053KB) ( 242 )   Save BACKGROUND: Interleukin-8 is an important inflammatory chemokine that plays an important role in the regulation of tumor cell proliferation and angiogenesis. OBJECTIVE: To investigate the effect of interleukin-8 on the invasion and metastasis of CD133+ hepatocellular carcinoma stem cells. METHODS: After isolation and culture of MHCC97-H cell lines, CD133+/CD133- MHCC97-H cells were sorted using immunomagnetic beads. CD133 expression was detected using flow cytometry, and interleukin-8 level in supernatant was measured using ELISA method. Cloning efficiency, tumorigenic capacity, cell migration and invasion ability were detected through colony formation assay, tumorigenesis experiment in nude mice, and Transwell detection. Additionally, other cells were neutralized using interleukin-8 neutralizing antibody. Measurement results were compared between cells undergoing different treatments. RESULTS AND CONCLUSION: The CD133 level, interleukin-8 level, cloning efficiency and cell membrane permeability of CD133+MHCC97-H cells were significantly higher than those of CD133-MHCC97-H cells (P < 0.05). Transplantation of CD133+MHCC97-H cells at 1×106/L and 1×107/L resulted in subcutaneous tumors in some mice, whereas no subcutaneous tumors appeared in mice undergoing transplantation of CD133-MHCC97-H cells at the same concentrations. After interleukin-8 neutralizing antibody treatment, the CD133 level, interleukin-8 level, and cloning efficiency of CD133+/CD133-MHCC97-H cells were significantly decreased (P < 0.05), especially in the CD133+MHCC97-H cells (P < 0.01); the migration and invasion ability and cell membrane permeability of CD133+MHCC97-H cells were significantly reduced (P < 0.05), but these changes were not obvious in CD133-MHCC97-H cells (P > 0.05). These results show that interleukin-8 could be specifically involved in the invasion and metastasis of CD133+MHCC97-H 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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Transplanting umbilical cord blood mesenchymal stem cells in ovarian cancer chemotherapy Li Xia, Wang Dong-hui, Guo Liang 2016, 20 (41):  6151-6157.  doi: 10.3969/j.issn.2095-4344.2016.41.011 Abstract ( 291 )   PDF (1022KB) ( 287 )   Save BACKGROUND: Existing evidence has confirmed that umbilical cord blood mesenchymal stem cells have an effect on functional recovery of a variety of damaged cells. OBJECTIVE: To explore the effect of umbilical cord blood mesenchymal stem cell transplantation on ovarian cancer chemotherapy. METHODS: Sixty healthy adult female Wistar rats were randomly divided into normal control group, damage group and treatment group (n=20/group). There was no treatment in the control group, and a rat model of ovarian cancer chemotherapy damage was made in the damage group and treatment group. After successful modeling, rats in the control group were given normal saline injection via the tail vein, and those in the damage and treatment groups were given paclitaxel chemotherapy and pacligaxel chemotherapy plus umbilical cord blood mesenchymal stem cell transplantation, respectively. After transplantation of 2 weeks, mRNA and protein expressions of XAF1 and Survivin in ovarian tumor tissues were detected by RT-PCR and western blot assay, respectively. Apoptosis in ovarian cancer cells were detected using TUNEL method. RESULTS AND CONCLUSION: Compared with the damage group, a significant up-regulation of XAF1 mRNA and protein but a remarkable down-regulation of Survivin mRNA and protein were obtained in the treatment group (P < 0.05). A severe damage to the ovarian tissues was visible in the damage group, presenting with large hemorrhage and necrosis area. This damage was markedly reduced in the treatment group. Additionally, the apoptotic rate of ovarian cancer cells was significantly higher in the treatment group than the damage group (P < 0.05). All these findings indicate that umbilical cord blood mesenchymal stem cell transplantation aids in ovarian cancer chemotherapy to promote ovarian tissue repair in rats, and XAF1 and Survivin cannot be ignored in tumor angiogenesis and ovarian cancer 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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Effect of transplantation of bone marrow mesenchymal stem cells on gastric cancer cell line SGC7901 in a rat model of gastric cancer Chen Chao, Li Hai-tao 2016, 20 (41):  6158-6163.  doi: 10.3969/j.issn.2095-4344.2016.41.012 Abstract ( 240 )   PDF (915KB) ( 282 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can migrate into tumor tissues, as reported in recent studies. OBJECTIVE: To investigate the tropism and effect of transplantation of bone marrow mesenchymal stem cells on proliferation and differentiation of gastric cancer cell line SGC7901 in rats. METHODS: Gastric cancer models were established in Sprague-Dawley rats by subcutaneous injection of gastric cancer cell line SGC7901, and then, in cell transplantation group, each rat underwent an intraperitoneal injection of 1×107 bone marrow mesenchymal stem cells. After transplantation, the targeting ability of bone marrow mesenchymal stem cells was detected using fluorescent DiI labeling. Cyclin D2 mRNA and protein expressions were measured using real-time PCR and western blot assay, respectively. Apoptosis of gastric cancer cells was observed by in situ terminal labeling method. RESULTS AND METHODS: Bone marrow mesenchymal stem cells successfully migrated into the gastric cancer site in rats. The expression levels of cyclin D2 mRNA and protein in the cell transplantation group were significantly higher than those in the model group (P < 0.05). It was observed that the number of apoptotic cancer cells was significantly reduced in the cell transplantation group compared with the model group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells have the ability to migrate into the tumor sites, thereby promoting the proliferation of gastric cancer 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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Sufentanil combined with umbilical cord blood stem cells protects against myocardial infarction Zhang Hong-yu 2016, 20 (41):  6164-6170.  doi: 10.3969/j.issn.2095-4344.2016.41.013 Abstract ( 267 )   PDF (1053KB) ( 227 )   Save BACKGROUND: In recent years, delayed protection of drug pretreatment has become a hot spot in the field of pretreatment, and sufentanil is a non-selective opioid receptor agonist that plays a significant myocardial protective role. OBJECTIVE: To investigate the protective effect of sufentanil combined with umbilical cord blood stem cell transplantation on injured myocardium after myocardial infarction. METHODS: Myocardial infarction model by ligating the left anterior descending coronary artery (ischemia 30 minutes and reperfusion 180 minutes) was made in 90 healthy adult Wistar rats, and then these rat models were randomly divided into ischemia-reperfusion (IR) group, cell transplantation group, combined treatment group (n=30/group). Rats in the IR group were given intraperitoneally intravenous injection of 1 mL normal saline 5 minutes before IR; rats in the cell transplantation group were given intraperitoneally intravenous injection of 1 mL umbilical cord blood stem cell suspension 5 minutes prior to IR; and rats in the combined treatment group were given intraperitoneally intravenous injection of 10 μg/kg sufentanil 10 minutes before IR and 1 mL umbilical cord blood stem cell suspension 5 minutes prior to IR. Two weeks later, related indicators were detected. RESULTS AND CONCLUSION: Compared with the IR group, myocardial infarct size was reduced significantly in the combined treatment group, followed by the cell transplantation group (P < 0.05). Lowest levels of serum creatine kinase, lactate dehydrogenase and troponin I were obtained in the combined treatment group followed by the cell transplantation group and IR group, while highest level of nitric oxide was observed in the combined treatment group followed by the cell transplantation group and IR group(P < 0.05). Highest expression of Caspase-3 protein was recorded in the combined treatment group, followed by the cell transplantation group and IR group (P < 0.05). Significantly increased left ventricular diastolic blood pressure and decreased left ventricular end diastolic pressure were found in the two cell transplantation groups compared with the IR group (P < 0.05). Moreover, pathological injury of the myocardial tissues was mitigated in the cell transplantation group and significantly relived in the combined treatment group compared with the IR group. In conclusion, the combined use of sufentanil and umbilical cord blood mesenchymal stem cell transplantation can reduce the degree of myocardial infarction and protect against myocardial 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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Adipose-derived mesenchymal stem cell transplantation for type 1 diabetes in a rat model Liu Xiao-hui 2016, 20 (41):  6171-6176.  doi: 10.3969/j.issn.2095-4344.2016.41.014 Abstract ( 243 )   PDF (993KB) ( 333 )   Save BACKGROUND: Stem cell transplantation is a promising treatment for type 1 diabetes mellitus. Adipose-derived mesenchymal stem cells have become another hotspot following bone marrow mesenchymal stem cells. OBJECTIVE: To investigate the therapeutic effect of adipose-derived mesenchymal stem cell transplantation on type 1 diabetes in a rat model. METHODS: Forty-five Sprague-Dawley rats were randomized into normal, model and cell transplantation. Animal model of type 1 diabetes was made in the latter two groups through intraperitoneal injection of streptozotocin. Seven days after modeling, rats in the three groups were given intraperitoneal injection of normal saline, serum-free DMEM or adipose-derived mesenchymal stem cell suspension, respectively. Two weeks after injection, body mass, blood glucose level, insulin secretion and PDX-1 mRNA in the pancreatic tissue of rats were monitored and detected in the three groups. RESULTS AND CONCLUSION: After modeling, the body mass of rats were lowered, and increased gradually in the cell transplantation group at 2 weeks after cell transplantation, but it was still decreased in the model group. Compared with the normal group, the fasting blood glucose level was significantly higher in the model group (P < 0.05), but it was reduced significantly after cell transplantation (P < 0.05). Compared with the normal group, the insulin level was reduced significantly in the model group (P < 0.05), but it was increased significantly after cell transplantation (P < 0.05). Highest and lowest PDX-1 mRNA expressions were obtained in the normal and model groups, respectively; and there was a significant difference between groups (P < 0.05). All these findings show that adipose-derived mesenchymal stem cell transplantation relieves hyperglycemia in rats by promoting the expression of PDX-1 in the rat pancreatic 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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Adipose-derived mesenchymal stem cell transplantation for treatment of chronic obstructive pulmonary disease Hao Shu-an, Wu Qi, Wang Mao-jun 2016, 20 (41):  6177-6182.  doi: 10.3969/j.issn.2095-4344.2016.41.015 Abstract ( 279 )   PDF (1039KB) ( 279 )   Save BACKGROUND: Chronic obstructive pulmonary disease is progressive respiratory disease characterized by airflow limitation. OBJECTIVE: To investigate the therapeutic effect of adipose-derived mesenchymal stem cell transplantation in rats with chronic obstructive pulmonary disease. METHODS: Sixty healthy male Sprague-Dawley rats were randomly divided into control, model and treatment groups (n=20 per group). Rat models of chronic obstructive pulmonary disease were made in the model and treatment group, while no treatment was done in the control group. After modeling, rats in the treatment group were given tail vein injection of CM-Dil-labeled adipose-derived mesenchymal stem cells, and rats in the model and control groups given the same amount of normal saline. Rat pulmonary ventilation function, inflammatory factor level and pathological changes of the lung were detected at 14 days after cell transplantation. RESULTS AND CONCLUSION: After modeling, reduced lung function was found in rats with chronic obstructive pulmonary disease, but the cell transplantation significantly improved this reduction (P < 0.05). Compared with the model group, significantly reduced indexes were visible in the treatment group (P < 0.05), including the total number of white blood cells, the number of macrophages and neutrophils, levels of interleukin-8, tumor necrosis factor α and C-reactive protein in the bronchoalveolar lavage fluid as well as serum level of malondialdehyde, while serum superoxide dismutase activity was increased significantly in the treatment group (P < 0.05). In the treatment group, emphysema-like changes were mitigated and CM-Dil-labeled adipose-derived mesenchymal stem cells were capable of homing to the lung tissue of rats with chronic obstructive pulmonary disease and survived. All these findings show that adipose-derived mesenchymal stem cell transplantation can improve lung function of rats with chronic obstructive pulmonary disease, and reduce inflammatory response, which has for certain some therapeutic effects.     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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Transplantation of erythropoietin gene-modified endothelial progenitor cells to treat lower extremity artery occlusion: a magnetically-labeled MRI evaluation Xu Guang-yu, Tian Su-hong, Zhou Shi-qi 2016, 20 (41):  6183-6189.  doi: 10.3969/j.issn.2095-4344.2016.41.016 Abstract ( 221 )   PDF (1170KB) ( 245 )   Save BACKGROUND: Erythropoietin and progenitor cell transplantation both have therapeutic effects on lower extremity arterial occlusive disease. OBJECTIVE: To investigate the erythropoietin modification effect and magnetic resonance imaging feasibility of superparamagnetic iron oxide (SPIO)-labeled endothelial progenitor cells in vitro. METHODS: Rat bone marrow-derived endothelial progenitor cells at logarithmic growth phase were randomized into four groups: endothelial progenitor cell group, SPIO labeled transfection group (pcDNA3-EPO transfection followed by SPIO labeling), SPIO labeled empty vector group (empty plasmid transfection followed by SPIO labeling), and SPIO labeling group (only SPIO labeling). 4.7T MRI was used to observe SPIO-labeled endothelial progenitor cells. Cell proliferation, cell cycle distribution, and expression of erythropoietin protein in the four groups were measured. RESULTS AND CONCLUSION: MRI findings showed with the increasing cell number, gradually lowered signal intensity on T1-weighted imaging (T1WI), T2WI and T2*WI was seen, and the reduction in the signal intensity was the maximum on T2*WI sequence and the minimum on T1WI sequence. For T1WI, T2WI and T2*WI sequences, the minimum number of cells was 2×104, 1×104 and 0.5×104, respectively. Cell proliferation and cell cycle distribution showed no significant difference among three SPIO labeling groups. In addition, the expression of erythropoietin protein was only found in the SPIO-labeled transfection group. These findings showed that under SPIO labeling, erythropoietin gene-modified endothelial progenitor cells show no changes in cell proliferation and cell cycle, and the 4.7T MR is capable of imaging SPIO-labeled erythropoietin gene-modified endothelial progenitor cells in vitro.     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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Cardiac stem cells: isolation, culture, proliferation and migration Hou Bo, Zhu Xian-yun, Wang Xue-kun 2016, 20 (41):  6190-6196.  doi: 10.3969/j.issn.2095-4344.2016.41.017 Abstract ( 352 )   PDF (1041KB) ( 257 )   Save BACKGROUND: In the process of cardiac stem cell culture in vitro, the growth microenvironment may have some effects on the cell proliferation. OBJECTIVE: To investigate the possible mechanism of proliferation and migration of rat cardiac stem cells cultured in vitro. METHODS: Cardiac tissues from 10 Sprague-Dawley rats were obtained for primary culture and subculture. Passage 3 cells were collected for immunofluorescence staining, and stem cell growth factor receptor (c-kit) and CD45, CD90 were detected. Cultured tissues were collected and randomly divided into two groups: in group 1, paraformaldehyde fixation, paraffin embedding, hematoxylin-eosin staining, Masson staining, and detecting apoptotic cells using TUNEL method were conducted; in group 2, EdU labeling of proliferated cells, immunofluorescent detection of c-kit positive expression, matrix metalloproteinases 2, 9 and transforming growth factor β1 using immunofluorescent staining were done. RESULTS AND CONCLUSION: After 7-10 days of myocardial tissue culture, bright and round cells were visible, and after adhesion, fusiform cells exhibited strong growth and proliferation ability. Immunofluorescence staining showed a large number of c-kit, CD45 positive cells but CD90 negative cells. After culture, a great number of newborn cells were found, accompanied by apoptosis of myocardial cells. After EdU staining, the positive cells were distributed in the myocardial gap, and showed a small amount of matrix metalloproteinases 2, 9 and transforming growth factor β1, while in the surrounding myocardium there was a large number of matrix metalloproteinases 2, 9 and transforming growth factor β1. Taken together, our findings show that cardiac stem cells could be obtained through myocardial tissue culture in vitro, accompanied by cell proliferation and migration. The mechanism is related to the expression of matrix metalloproteinases 2, 9 and transforming growth factor β1.     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 of adipose-derived stem cells derived from renal fat capsule and groin in vitro Zhu Yong-sheng, Deng Qing-fu, Li Jun 2016, 20 (41):  6197-6202.  doi: 10.3969/j.issn.2095-4344.2016.41.018 Abstract ( 330 )   PDF (3645KB) ( 222 )   Save BACKGROUND: Adipose-derived stem cells have different sources, but it is unclear whether these cells from different sources have difference in their biological properties. OBJECTIVE: To detect the in vitro proliferation and chondrogenic differentiation of adipose-derived stem cells derived from renal fat capsule and groin. METHODS: Adipose-derived stem cells from renal fat capsule and groin of rats were isolated, cultured and identified. MTT assay was used to detect in vitro proliferation ability of these cells. Passage 3 cells were under chondrogenic induction for 2 weeks. After induction, the expression of type II collagen was observed by immunofluorescence detection, and RT-PCR was employed to detect the expression levels of Aggrecan and type II collagen mRNA in the two groups. RESULTS AND CONCLUSION: After primary culture and passage, adipose-derived stem cells from the renal fat capsule and groin of rats exhibited similar morphology, and over 95% of cells expressed CD44 in the two groups. Adipose-derived stem cells from two sources showed an S-shaped growth curve in vitro and were positive for type II collagen. After RT-PCR detection, the expression levels of Aggrecan and type II collagen mRNA had no difference in adipose-derived stem cells from renal fat capsule and groin (P > 0.05). Experimental results show that adipose-derived stem cells from both renal fat capsule and groin exhibit stable growth, rapid proliferation and chondrogenic differentiation under orient induction in vitro, indicating there is no difference between these cells from two sources.     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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Overexpression of glycogen synthase kinase 3beta in the treatment of myocardial infarction with cardiac stem cell transplantation Zhao Cui-hua, Li Yan-ming, Zhong Xiao-ming, He Rui-li, Cheng Guan-chang 2016, 20 (41):  6203-6208.  doi: 10.3969/j.issn.2095-4344.2016.41.019 Abstract ( 226 )   PDF (4088KB) ( 272 )   Save BACKGROUND: The mechanism and effect of glycogen synthase kinase 3β (GSK-3β) in the differentiation of cardiac stem cells into cardiomyocytes are still unclear, although GSK-3β is closely related to the life activities of cells. OBJECTIVE: To investigate the changes of GSK-3β expression in the treatment of myocardial infarction in rats undergoing cardiac stem cell transplantation. METHODS: The isolation and culture of cardiac stem cells were performed in 10 neonatal rats. Lentivirus overexpressing GSK-3β or LacZ (control) was constructed and transferred into cardiac stem cells. Animal model of myocardial infarction was made in 30 Sprague-Dawley rats. Six weeks after model preparation, rat models were assigned into GSK-3β, LacZ or PBS group. GSK-3β or LacZ overexpressing cardiac stem cell solution or PBS in equal volume was injected into the rat myocardium, respectively. Four weeks after transplantation, the cardiac function and myocardial collagen production in rats were detected and compared. RESULTS AND CONCLUSION: Compared with the other two groups, the left ventricular ejection fraction was significantly higher, and the left ventricular end diastolic diameter was significantly lower in the GSK-3β group (P < 0.05). Hydroxyproline content, type I collagen mRNA, and type III collagen mRNA expression were significantly lower in the GSK-3β group than the other two groups (P < 0.05). Findings from Masson staining showed that the content of blue-stained collagen was significantly lower in the GSK-3β group than the LacZ group. Moreover, lowest myocardial infarction size was found in the GSK-3β group (P < 0.05). All these experimental findings show that GSK-3 overexpression plays a positive role in promoting the therapeutic effect of cardiac stem cell transplantation.     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 effect of exosomes in mesenchymal stem cell differentiation Hou Xiao-can, Jia Yan-jie, Peng Tao 2016, 20 (41):  6209-6215.  doi: 10.3969/j.issn.2095-4344.2016.41.020 Abstract ( 323 )   PDF (981KB) ( 354 )   Save BACKGROUND: Currently, what we know about exosomes is that it can be produced by a variety of cells and transfer a variety of materials, producing subsequent function of regulation. OBJECTIVE: To review the function of exosomes in mesenchymal stem cell differentiation, in order to provide reference for further in-depth study. METHODS: With the key words of “exosome, mesenchymal stem cell, differentiation” in Chinese and English, respectively, a computer-based search was performed for articles published in CNKI, Medline and Embase databases from January 2001 to September 2016. After the initial screening, the reserved articles were further detailed, summarized and concluded. RESULTS AND CONCLUSION: Exosomes are a kind of vesicles 40-100 nm in diameter, which can be secreted by a variety of cell types, containing functional products, such as functional protein, gene product, lipid and so on. As a bridge of adjacent cells transferring functional products, exosomes are becoming an issue of concern in the microenvironment for cell interaction. In the differentiation of mesenchymal stem cells, exosomes also play an indispensable role via pathway regulation.     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 in immunomodulatory properties of mesenchymal stem cells Yi Qiao, Lu Yan-qin, Huang Hong-yu, Zhou Dian, Liu Ou-sheng 2016, 20 (41):  6216-6224.  doi: 10.3969/j.issn.2095-4344.2016.41.021 Abstract ( 358 )   PDF (1174KB) ( 270 )   Save BACKGROUND: Mesenchymal stem cells are multipotent progenitor cells that can be isolated from the bone marrow, adipose tissue, umbilical cord blood and so on. Mesenchymal stem cells possess strong immunosuppressive effects on both innate and adaptive immunity. OBJECTIVE: To summarize the immunomodulatory properties of mesenchymal stem cells for T lymphocytes, B lymphocytes, natural killer cells and dentritic cells and prospect its therapeutic implication. METHODS: PubMed and CNKI databases were searched for relevant literatures published from 2000 to 2015. The key words were “mesenchymal stem cells, immunomodulator, T cells, B cells, NK cells, dendriticc cells” in Chinese and English, respectively. Then, 61 papers were included and further analyzed. RESULTS AND CONCLUSION: The immunomodulatory effects of MSCs on lymphocytes, B lymphocytes, natural killer cells and dentritic cells were elicited by cell-cell contact and soluble cytokines. But mesenchymal stem cells from different sources hold different immunomodulatory effect on immune cells. When we use mesenchymal stem cells in clinic, some important factors, such as isolation methods, cell sources, colonization sites, should be taken into account. In order to ensure the clinical safety and effectiveness, there are still many problems to be further studied before mesenchymal stem cells can be widely used in clinic.     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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Recent advances in surface markers of gastric cancer stem cells Du Hua, Shi Ying-xu 2016, 20 (41):  6225-6232.  doi: 10.3969/j.issn.2095-4344.2016.41.022 Abstract ( 336 )   PDF (997KB) ( 279 )   Save BACKGROUND: It has been observed recently that tumor recurrence is closely related to cancer stem cells. Cancer stem cell theory provides a new avenue for the study of gastric cancer pathogenesis, diagnosis and treatment. To determine the cell phenotype is helpful to target gastric cancer stem cells, and contributes to exploring the self-renewal and differentiation of gastric cancer stem cells, further providing new ideas for the treatment of gastric cancer. However, the phenotype of gastric cancer stem cells remains controversial. OBJECTIVE: To review the research progress in gastric cancer stem cell phenotype. METHODS: A computer-based online search of PubMed database was performed to search related articles published between January 1965 and October 2015 with the key words of “Gastric Cancer Stem Cells” in English. Sixty-eight articles were included for final analysis. RESULTS AND CONCLUSION: Traditional cancer stem cell phenotypes are unsuitable for marking gastric cancer stem cells. Studies have found that CD90 expresses in primary gastric cancers, and cancer stem cell spheres would be required via serum-free culture by CD90 maker. CD24 expression can improve the adhesion, invasion and migration of gastric cancer cells. Further studies have confirmed that a lot of cells positive for both CD44 and CD54 can be found in patients with early recurrence of gastric cancer rather than those with advanced recurrence, and CD44 and CD54 expression in tumor cells is likely to be an important cause of gastric cancer occurrence and recurrence. By analyzing the sorting, CD44+CD24+CD90+CD54+ is likely to be the gastric cancer stem cell 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