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04 June 2015, Volume 19 Issue 23 Previous Issue    Next Issue

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Bone tissue regeneration of bone morphogenetic protein-7-transfected bone marrow mesenchymal stem cells  Wang Ping, Gu Yu-cen, Gao Zhi-hui, Sun Tie-feng, Yang Wu-bin 2015, 19 (23):  3609-3615.  doi: 10.3969/j.issn.2095-4344.2015.23.001 Abstract ( 277 )   PDF (1357KB) ( 550 )   Save BACKGROUND: Bone morphogenetic protein-7 can promote proliferation of bone marrow mesenchymal stem cells and has important significance in terms of inducing new bone formation. OBJECTIVE: To observe the protein and mRNA expression of bone morphogenetic protein-7 in bone marrow mesenchymal stem cells transfected with bone morphogenetic protein-7. METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured, and then identified by the morphology and cell surface markers. Lentiviral vectors carrying bone morphogenetic protein-7 were constructed to transfect bone marrow mesenchymal stem cells. After total RNA extraction, the protein and mRNA expression of bone morphogenetic protein-7 was detected by RT-PCR and western blot methods. RESULTS AND CONCLUSION: PLV-sfGFP(2A)-BMP7 recombinant plasmids were successfully constructed and transferred into rat bone marrow mesenchymal stem cells. The mRNA and protein expression of bone morphogenetic protein-7 in the transfection group was significantly higher than that in the blank vector and blank groups (P < 0.05), indicating exogenous bone morphogenetic protein-7 gene is effectively integrated into the 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 p53 inhibitor on viability of human bone marrow mesenchymal stem cells in late-phase amplification  He Ze-bin, Zhao Yun-he, Yang Gui-jiao, Lu Li 2015, 19 (23):  3616-3620.  doi: 10.3969/j.issn.2095-4344.2015.23.002 Abstract ( 319 )   PDF (943KB) ( 336 )   Save BACKGROUND: It is not fully understood that whether p53 inhibitor can directly intervene in the viability of bone marrow mesenchymal stem cells and the possible mechanism. OBJECTIVE: To investigate the effect of p53 inhibitor, PFT-α, on the aging process of bone marrow mesenchymal stem cells in late-phase amplification and to discover the key target to delay the replicative senescence of human bone marrow mesenchymal stem cells. METHODS: The expression levels of p53, p21, and p15 mRNA in human bone marrow mesenchymal stem cells in both early and late-phase amplification were detected by quantitative PCR assay. Then, human bone marrow mesenchymal stem cells in late-phase amplification were respectively treated with 20 µmol/L PFT-α or an equivalent amount of dimethyl sulfoxide for 2 weeks. The positive rate of aging cells was determined by SA-β-Gal staining. The apoptosis was detected by TUNEL staining. Human bone marrow mesenchymal stem cells were treated with 300 µmol/L H2O2 for 30 minutes, and then cellular anti-oxidative stress capacity was detected by cell counting kit-8 assay. RESULTS AND CONCLUSION: The quantitative PCR assay showed that the mRNA expression level of p15,  p21 and p53 in human bone marrow mesenchymal stem cells in late-phase amplification was significantly increased (1.45±0.23), (1.51±0.14) and (1.78±0.14) times as much as that in early phase amplification (P < 0.05). The positive rate of aging cells in PFT-α group was significantly lower than that in the dimethyl sulfoxide group [(41±5)% vs. (63±7)%, P < 0.05)]. However, there was no significant difference in apoptosis rate between PFT-α group and dimethyl sulfoxide group. After treatment with H2O2, the absorbance value in the PFT-α group was (1.27±0.13) times as much as that in the dimethyl sulfoxide group (P < 0.001). The above results demonstrate that the activation of p53 signaling pathway may be an important factor of causing aging of human bone marrow mesenchymal stem cells. Application of p53 inhibitor PFT-α can enhance the anti-oxidative stress capacity of human bone marrow mesenchymal stem cells in late phrase amplification. 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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Construction of human telomerase reverse transcriptase-immortalized rat bone marrow mesenchemal stem cell strains  Liu Hui-ping, Zhong Xiao-long, Zhao Qing, Huang Wen-qi, An Ke 2015, 19 (23):  3621-3627.  doi: 10.3969/j.issn.2095-4344.2015.23.003 Abstract ( 400 )   PDF (5480KB) ( 585 )   Save BACKGROUND: Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cells are the ideal cell type to serve as vectors of transgenic cells in pain management. However, the replicative senescence and small amount of cells obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cells in pain research. OBJECTIVE: To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cells as transgenic cellular vectors for pain therapy. METHODS: Bone marrow mesenchymal stem cells were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) followed by puromycin selection. hTERT expression and telomerase activity in these transfected cells were determined by RT-PCR and TRAP. Morphological changes, capacity of cell growth and multi-lineage differentiation, chromosome karyotype and  tumorigenicity were observed in vitro. Moreover, the expression of cell surface molecule, Nestin, MHC-I and MHC-II in transfected cells were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cells genetically modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cells, the hTERT-modified bone marrow mesenchymal stem cells showed higher expression of hTERT mRNA, telomerase activity and cell proliferation. Most of transfected cells stayed at G2/M and S stages. The proliferation index of the transfected cells were increased dramatically. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cells were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cell line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cells, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cells. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cells immortalized by hTERT gene are constructed successfully and still maintain major stem cells characteristics, which provide safe and stable cell vectors as research base for pain therapy. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells with reduced glutathione against mouse lung injury  Chen Fei, Li Guo-qing, Hu Feng-qing, Mei Ju, Wang Ming-song 2015, 19 (23):  3628-3632.  doi: 10.3969/j.issn.2095-4344.2015.23.004 Abstract ( 297 )   PDF (4715KB) ( 276 )   Save BACKGROUND: A large number of experiments have shown that bone marrow mesenchymal stem cells (BMSCs) have good effects on the treatment of lung disease or improvement of lung injury, and its therapeutic effect is mainly oriented to reduce the inflammatory response. OBJECTIVE: To study the effects of BMSCs combined with reduced glutathione in murine models of bleomycin-induced lung injury. METHODS: BMSCs were isolated from a male NOD/SCID mouse and cell morphology and phenotype were  observed. Sixty-four female NOD/SCID mice were randomly divided into a control group, a model group, a BMSCs group and a BMSCs+reduced glutathione group (combined group). The control group received intratracheal injection of normal saline, the model group received intratracheal injection of bleomycin, and the BMSC group received BMSCs injection via the tail vein at 2 hours after intratracheal injection of bleomycin, and combined group received BMSCs+reduced glutathione injection via the tail vein at 2 hours after intratracheal injection of bleomycin. All mice were killed after 7 days, and levels of tumor necrosis factor-α, interleukin-β, malondialdehyde in lung tissue were detected and lung tissue specimens were obtained for pathological examination. RESULTS AND CONCLUSION: BMSCs were fibroblast-like cells, Positive for CD10, CD13 and CD44, but negative for CD34 and CD45. Compared with the control group, the levels of tumor necrosis factor-α, interleukin-β and malondialdehyde were increased in the model group, while decreased significantly in the BMSCs group and combined group (P < 0.05), especially in the combined group (P < 0.05). Pathological examination showed that more serious lung injury was observed in the model group and BMSCs group than the combined group. These findings indicate that BMSCs combined with reduced glutathione can more effectively protect against bleomycin-induced lung 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 Ginsenoside Rh2 on oxytocin-induced transformation of bone marrow mesenchymal stem cells into myocardial cells  Wang Le, Tian Li, Zheng Ming-qi, Liu Gang, Ji Li-shuang, Ma Guo-ping 2015, 19 (23):  3633-3638.  doi: 10.3969/j.issn.2095-4344.2015.23.005 Abstract ( 250 )   PDF (4733KB) ( 262 )   Save BACKGROUND: Our prior experiments have confirmed that 10 μmol/L oxytocin can induce transformation of bone marrow mesenchymal stem cells into myocardial cells. OBJECTIVE: To investigate the effects of Ginsenoside Rh2 on oxytocin-induced transformation of bone marrow mesenchymal stem cells into myocardial cells. METHODS: Rat bone marrow mesenchymal stem cells were isolated by differential adherence method. These isolated cells were randomly divided into five groups. In the blank control group, cells were routinely cultured. In the oxytocin group, cells were cultured with 10 μmol/L oxytocin for 2 consecutive weeks. In the Ginsenoside Rh2 low-, middle-, and high-dose groups, cells were treated with 0.5, 1, 2 μmol/L Ginsenoside Rh2 respectively for 24 hours and then with oxytocin for additional 2 consecutive weeks. RESULTS AND CONCLUSION: Optical microscopy showed that compared to the blank control group, some cells in the oxytocin group exhibited an increased soma and some cells grew in clusters and the cell clusters enlarged with the increase in Ginsenoside Rh2 dose. Immunocytochemical staining and western blot analysis showed that cardiac Troponin T and connexin 43 protein expression in the oxytocin, Ginsenoside Rh2 low-, middle-, and high-dose groups were significantly greater than in the blank control group (P < 0.05), and cardiac Troponin T and connexin 43 protein expression in the Ginsenoside Rh2 groups was increased with the increase  in Ginsenoside Rh2 dose and was significantly higher than that in the oxytocin group (P < 0.05). Confocal laser scanning microscopy showed that the relative fluorescence intensity of free calcium in the bone marrow mesenchymal stem cells in the oxytocin group was significantly increased after induction by oxytocin for 2 weeks (P < 0.05), while the relative fluorescence intensity in the Ginsenoside Rh2 groups was significantly higher than that in the Ginsenoside Rh2 groups and was positively correlated with the dose of Ginsenoside Rh2. These findings suggest that Ginsenoside Rh2 can obviously promote oxytocin-induced transformation of bone marrow mesenchymal stem cells into myocardial 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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The mechanism by which bone marrow mesenchymal stem cells participate in apoptosis of hepatic stellate cells Liu Da-li 2015, 19 (23):  3639-3643.  doi: 10.3969/j.issn.2095-4344.2015.23.006 Abstract ( 260 )   PDF (3864KB) ( 303 )   Save BACKGROUND: Control of hepatic stellate cell activation and proliferation is the focus of developing strategies against liver fibrosis. Human or murine bone marrow mesenchymal stem cells can induce apoptosis of hepatic stellate cells through paracrine of hepatocyte growth factors. OBJECTIVE: To explore the mechanism by which bone marrow mesenchymal stem cells participate in apoptosis of rat hepatic stellate cells. METHODS: Hepatic stellate cells and bone marrow mesenchymal stem cells were seeded and co-cultured in the upper and lower chambers in a co-culture system, serving as a co-culture group. In the blank control group, only hepatic stellate cells were involved. In the c-Met inhibitor group, hepatic stellate cells and bone marrow mesenchymal stem cells were treated with 3 mg/L C-Met inhibitor. In the RhoA inhibitor group, both kinds of cells were treated with 3 mg/L RhoA inhibitor. RESULTS AND CONCLUSION: The concentration of c-Met inhibitor was 3.0 mg/L. RhoA inhibitor at 30 μmol/L exhibited a greater inhibitory effect than at other concentrations. RhoA mRNA and protein expression in the co-culture, c-Met inhibitor and in particular RhoA inhibitor groups was obviously greater than in the blank control group. Hepatocyte growth factor concentration in each group was gradually decreased with time, hepatocyte growth factor activator concentration in each group was gradually increased with time, and the changes were most obvious in the c-Met inhibitor group. Apoptosis rate of hepatic stellate cells in each group was gradually increased with time, and highest apoptosis rate appeared in the RhoA inhibitor group, and lowest apoptosis rate in the c-Met inhibitor group. These findings suggest that bone marrow mesenchymal stem cells participate in and promote the apoptosis of hepatic stellate cells by activating hepatocyte growth factors and downregulating Rho activity. 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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Serum from liver injury rats induces differentiation of umbilical cord mesenchymal stem cells into hepatocyte-like cells   Zhong Yan, Tang Xiao-peng 2015, 19 (23):  3644-3651.  doi: 10.3969/j.issn.2095-4344.2015.23.007 Abstract ( 258 )   PDF (7849KB) ( 305 )   Save BACKGROUND: Umbilical cord mesenchymal stem cells are from the umbilical cord of newly born individuals and have no ethical issues, and therefore are promising candidates for seeded cells as a substitute for cell transplantation and regenerative medicine. OBJECTIVE: To investigate the effects of serum from liver injury rats on induced differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells and provide experimental evidence for use of human umbilical cord mesenchymal stem cells in the treatment of patients with end-stage liver disease in the clinic. METHODS: Rat models of acute liver injury were established by intraperitoneal injection of 10% carbon tetrachloride. Rats in the control group were intraperitoneally administered the same amount of soybean oil. Forty-eight hours after modeling, abdominal aorta blood was taken for serum preparation. Passage 3 human umbilical cord mesenchymal stem cells were cultured with 20% serum from liver injury rats and 20% fetal bovine serum. Morphology of human umbilical cord mesenchymal stem cells was observed before and after culture. Levels of α-fetoprotein and albumin in the supernatant were detected. RESULTS AND CONCLUSION: Cells exhibited shuttle-shaped appearance and grew in whirlpool-like manner at 1 day after culture with serum from liver injury rats, exhibited short shuttle-shaped appearance at 2 days, were oval-shaped at 3 days, and were round and an extremely small number of cells were floated at 4 days. At 4 days after culture with serum from liver injury rats, level of albumin in the cell supernatant was significantly increased than that before induction and that in the control group (P < 0.001), and there was no significant difference in level of α-fetoprotein in the cell supernatant. These results suggest that serum of liver injury rats can induce differentiation of umbilical cord mesenchymal stem cells into hepatocyte-like 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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Differentiation of bone marrow and umbilical cord blood mesenchymal stem cells into osteoblasts in vitro: comparison of their osteogenic potentials  Shao Shuai, Zhou Chen-hong, Xu Li-li 2015, 19 (23):  3652-3657.  doi: 10.3969/j.issn.2095-4344.2015.23.008 Abstract ( 291 )   PDF (8572KB) ( 747 )   Save BACKGROUND: Mesenchymal stem cells isolated from cord blood and bone marrow have multi-directional differentiation ability under a certain condition of induction. OBJECTIVE: To compare the difference of differentiation of umbilical cord blood and bone marrow mesenchymal stem cells into osteoblasts. METHODS: Human umbilical cord blood and bone marrow mesenchymal stem cells were isolated and cultured by density gradient method. When reached 90% confluency, mesenchymal stem cells were digested by trypsin for subculture. At the third passage, umbilical cord blood mesenchymal stem cells and bone marrow mesenchymal stem cells at 8×104/well were incubated. When reached 80% confluency, cells were treated with low-glucose DMEM supplemented with 10% fetal bovine serum, 0.1 μmol/L dexamethasone, 50 μmol/L vitamin C and 10 mmol/L β-sodium glycerophosphate. RESULTS AND CONCLUSION: There was no significant difference in morphology and biological properties of the two kinds of mesenchymal stem cells. Cells were highly expressed CD44, CD29, but did not express CD34. They had the ability to differentiate into osteoblasts, which had a positive staining for known markers: alkaline phospatase and calcium in vitro mineralization. There was no significant difference in the activity of osteoblasts of two kinds of cells. Results verify that umbilical cord blood and adult bone marrow mesenchymal stem cells can be induced into osteoblasts with a similar ability, and they can be used as seed cells for bone tissue engineering.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord blood-derived CD133 cells for treatment of gestational diabetes  Ning Li, Yuan Yu, Tan Bin 2015, 19 (23):  3658-3663.  doi: 10.3969/j.issn.2095-4344.2015.23.009 Abstract ( 285 )   PDF (4779KB) ( 246 )   Save BACKGROUND: Human umbilical cord blood contains a large number of hemopoietic stem cells and mesenchymal stem cells. Neonate umbilical cord blood-derived stem cells show milder immunological rejection and lower immunogenicity than peripheral blood- and bone marrow-derived mesenchymal stem cells. OBJECTIVE: To investigate the feasibility of umbilical cord blood-derived CD133 cells for treatment of gestational diabetes, so as to provide novel methods for clinical treatment of gestational diabetes. METHODS: Mouse models of gestational diabetes were established by high-fat and high-sugar diet feeding. Umbilical cord blood-derived CD133 cells were sorted by flow cytometry sorting technique and then transplanted into mouse models of gestational diabetes via the tail vein. At 7 days after cell transplantation, serum levels of fasting blood glucose, fasting insulin, total cholesterol and triacylglycerol were measured and mouse pancreatic tissue injury and repair was observed by hematoxylin-esoin staining. Mouse insulin resistance index and pancreatic islet function were analyzed using the homeostatic model assessment. RESULTS AND CONCLUSION: Umbilical cord blood-derived CD133 cells could be isolated from umbilical cord blood monocytes using flow cytometry sorting technique, with cell purity of (90.24±2.56)%. At 7 days after cell transplantation, serum levels of fasting blood glucose, fasting insulin, total cholesterol and triacylglycerol were  significantly decreased, insulin resistance index was significantly decreased, and pancreatic islet function was significantly improved in mouse models of gestational diabetes (all P < 0.05). In addition, atrophy of pancreatic tissue and infiltration of inflammatory cells were reduced. These results show that umbilical cord blood-derived CD133 cell transplantation can improve the disease condition of gestational diabetes by repairing injured pancreatic tissue, decreasing insulin resistance index and improving pancreatic islet function.  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 different preservation solutions and placing time on survival rate of umbilical cord-derived mesenchymal stem cells Li Zhi-gang, Ben Liang, Han Yu, Bi Wei-wei 2015, 19 (23):  3664-3668.  doi: 10.3969/j.issn.2095-4344.2015.23.010 Abstract ( 510 )   PDF (4905KB) ( 391 )   Save BACKGROUND: The relationship between placing time after stem cell preparation and cell survival is the basis of safety and effectiveness for the clinical application. OBJECTIVE: To observe effects of different preservation solutions and different storage time on survival rate of umbilical cord-derived mesenchymal stem cells, and to provide important evidence for identifying effectiveness of stem cells. METHODS: Umbilical cord-derived mesenchymal stem cells were selected to prepare stem cell preparation, which was preserved in physiological saline, medium, medium+physiological saline, physiological saline containing epidermal growth factor, and medium containing low molecular heparin calcium suspension. Cold closet was selected for imitating cellular transport conditions. Samples were obtained at 0, 6, 12, 18, 24, 30, 36, 42 and 48 hours. Total cell number and cell survival rate were detected. RESULTS AND CONCLUSION: The difference in cell number and survival rate was not great within 24 hours in each group. Twenty-four hours later, total cell number and survival rate were better in the medium and medium containing low molecular heparin calcium groups than in the physiological saline containing epidermal growth factor, physiological saline, and medium+physiological saline groups. These findings suggest that after stem cell preparation, the cell survival rate can reach more than 90% within 24 hours under refrigerated transport conditions. Nutritional ingredients and proper pH value of preservation solution can make the cell survival rate increased greatly.  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 cancer stem cell surface marker CD44 in gastric cancer invasion and lymph node metastasis  Li Shi-ting, Tao Wen-cheng 2015, 19 (23):  3669-3673.  doi: 10.3969/j.issn.2095-4344.2015.23.011 Abstract ( 348 )   PDF (4101KB) ( 343 )   Save BACKGROUND: Tumor stem cells not only initiate tumorigenesis, but also are involved in the invasion and metastasis of tumor cells. For tumor stem cells, to identify the specific cell surface marker has become a research hotspot. OBJECTIVE: To explore the clinical significance of cancer stem cell surface marker CD44 in gastric cancer invasion and lymph node metastasis. METHODS: CD44 protein expression in specimens of gastric cancer tissue was detected by the immunohistochemical SABC method. The relationship between CD44 protein expression and biological characteristics and prognosis of gastric cancer was detected using Pearson χ2 test and Cox regression analysis. RESULTS AND CONCLUSION: Among 100 cases of gastric carcinoma, 59 cases (59%) were positive for CD44 protein expression. CD44 protein expression in normal gastric mucosa at above 5 cm from the edge of primary gastric cancer was negative. CD44 protein was widely expressed in tissues of gastric cancer, mainly expressed in the cell membrane, and a small amount of expression in the cytoplasm. CD44 protein expression in gastric cancer tissue was not correlated with sex of the patients or age (P > 0.05), but was associated with tumor staging and lymph duct tissue infiltration, histological grade, and tumor size (P < 0.05). Deep tumor  invasion, high histological grade, big diameter of tumor, and lymph node metastasis could lead to high positive CD44 protein expression. Positive expression of CD44 is an independent prognostic factor affecting postoperative survival (P < 0.05). The results show that the cancer stem cell surface markers CD44 in gastric carcinoma tissues is strongly associated with invasion of gastric carcinoma and lymph node metastasis. High expression of CD44 presents poor prognosis. 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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MicroRNA differential expression in liver cirrhosis rats undergoing human umbilical cord mesenchymal stem cell transplantation  Liu Xiang-zhong, Zou Zhi-qiang, Wang Gui-qiang, Li Dong, Shao Zhi-ying 2015, 19 (23):  3674-3680.  doi: 10.3969/j.issn.2095-4344.2015.23.012 Abstract ( 264 )   PDF (6686KB) ( 343 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) can obviously relieve liver cirrhosis, and thereby repair liver injury. However, the molecular mechanism of hUC-MSCs therapy for liver cirrhosis is limited at present, and especially the non-coding RNA regulation of hepatic gene changes has not been detailed. OBJECTIVE: To investigate the changes of microRNA after hUC-MSCs therapy in rats with liver cirrhosis. METHODS: Liver cirrhosis models were established in rats using carbon tetrachloride subcutaneous injection  plus oral administration of alcohol. At 8 weeks after modeling, hUC-MSCs were injected via the tail vein once a week for 4 consecutive weeks. At 1 week after the last injection, rat liver tissues were collected for paraffin embedding. Liver RNA was extracted for gene chip analysis. Blood samples were collected and analyzed using an automatic biochemical analyzer to detect the changes of liver function. RESULTS AND CONCLUSION: Alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl transpeptidase were improved significantly after hUC-MSCs therapy. Fat lesions and necrosis of hepatocytes were significantly reduced. MicroRNA expression microarray hybridization analysis and PCR results showed that rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153* were down-regulated during modeling and increased after hUC-MSCs therapy. And rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a were firstly up-regulated in the process of modeling and then down-regulated obviously after hUC-MSCs therapy. These results suggest that hUC-MSCs may reverse liver cirrhosis and liver cell damage through up-regulation of rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153*, and down-regulation of rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a.   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 | Related Articles | Metrics

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Spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells  Hao Guang-jun, Liu Qing, Wang Juan, Ma Yan-e1 Ding Yan-hui 2015, 19 (23):  3681-3685.  doi: 10.3969/j.issn.2095-4344.2015.23.013 Abstract ( 383 )   PDF (891KB) ( 286 )   Save BACKGROUND: There is no clear conclusion on whether the lung cancer stem cells can induce to spheroid formation and have tumorigenicity. OBJECTIVE: To observe the spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells. METHODS: SPC-A1 at proliferating phase was cultured in serum-free DF12 culture medium, and then recombinant human insulin-like growth factor-1, recombinant human epidermal growth factor, and recombinant human fibroblast growth factor-10 were added to induce spheroid cells. Immune fluorescence detection and PCR amplification were done to understand the expression of stem cell associated markers. NOD-SCID immunodeficient mice were subcutaneously implanted with lung spheroid cells to observe the tumor growth. In vivo fluorescence imager was used for radiography. RESULTS AND CONCLUSION: After 5-10 days, lung spheroid cells were harvested. RT-PCR results showed that lung spheroid cells were positive for CD24, CD221, CCSP and SP-C. In addition, the lung spheroid cells and purified CD24+, CD221+ lung cancer stem cells were both positive for TTF-1 of lung stem cells, OCT4 and Nanog of embryonic stem cells and TTF-1 of Bmi-1 lung stem cells. The fluorescence detection showed that over 80% lung spheroid cells expressed CCSP and OCT4; SPC-A1 cells had the characteristics of alveolar type II cells, and also expressed SP-C protein, but only about 5% of the cells expressed CCSP and OCT4. At 50 days after subcutaneous implantation of lung spheroid cells, in vivo fluorescence imaging showed that the diameter of tumor  in mice was 1 cm, indicating human lung adenocarcinoma cell line SPC-A1 can induce the spheroid formation, and lung cancer stem cells rich in the cell spheres have the tumorigenic ability. 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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Human telomerase reverse transcriptase gene-modified umbilical cord mesenchymal stem cell transplantation for acute kidney injury   Wang Tian-sheng, Zhang Jian-jun 2015, 19 (23):  3686-3691.  doi: 10.3969/j.issn.2095-4344.2015.23.014 Abstract ( 276 )   PDF (1166KB) ( 327 )   Save BACKGROUND: The human telomerase reverse transcriptase (hTERT) is one of preferred growth factors for regulating proliferation and directional differentiation, has multiple biological effects, and laids the foundation for genetically engineered immortalized stem cell lines. OBJECTIVE: To investigate the effect of hTERT gene-modified umbilical cord mesenchymal stem cell transplantation on acute kidney injury induced by ischemia and reperfusion in rats. METHODS: The human umbilical cord mesenchymal stem cells were cultured in vitro. Rat models of acute kidney injury induced by ischemia and reperfusion were established. Rat models were randomly divided into three groups. Rats in the control group were injected with 1 mL L-DMEM medium through caudal vein. Rats in the negative transfection group were injected with 1 mL umbilical cord mesenchymal stem cell suspension after empty virus transfection through caudal vein. Rats in the hTERT transfection group were injected with 1 mL umbilical cord mesenchymal stem cell suspension after PLXSN-hTERT transfection through caudal vein. RESULTS AND CONCLUSION: At 3 and 28 days after transplantation, hematoxylin-eosin staining showed renal tubular damage score in the hTERT transfection group < negative transfection group < control group (P < 0.05). At 28 days after transplantation, the number of CM-Dil-positive cells in the hTERT transfection group > negative transfection group > control group (P < 0.05). At 1, 3, 14, and 28 days, serum creatinine and urea nitrogen levels  in the hTERT transfection group < negative transfection group < control group (P < 0.05). The results confirm that hTERT gene-modified umbilical cord mesenchymal stem cell transplantation has a significant repair effect on acute kidney injury in rats.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Autologous bone marrow stem cell transplantation for thromboangiitis obliterans: 5-year follow-up Bai Chao, Guo Chen-ming, Luo Jun 2015, 19 (23):  3692-3697.  doi: 10.3969/j.issn.2095-4344.2015.23.015 Abstract ( 362 )   PDF (1053KB) ( 302 )   Save BACKGROUND: The assessment for long-term efficacy of chronic ischemic disease is more important than the short-term efficacy assessment, which associates with patient’s long-term quality of life and long-term survival rate. OBJECTIVE: To observe the 5-year follow-up outcomes of autologous bone marrow stem cell transplantation for the treatment of thromboangiitis obliterans. METHODS: This study enrolled 43 patients of thromboangiitis obliterans who underwent autologous bone marrow stem cell transplantation from August 2007 to January 2010 in the Department of Thyroid Vascular Surgery, the First Affiliated Hospital of Xinjiang Medical University. At 1, 2, 3, 4 and 5 years after transplantation, pain, cold sensation, and intermittent claudication distance were followed up by telephone; changes in limb ulcers were observed. At 1 year after transplantation, venous oxygen partial pressure and oxygen saturation of limbs were reviewed. RESULTS AND CONCLUSION: A total of 38 thromboangiitis obliterans patients with complete follow-up data were included in the final analysis. Compared to the preoperation, pain, cold sensation, and intermittent  claudication significantly improved. The difference was statistically significant (Z values: -4.277, -5.086, -3.574, P < 0.001). Compared with 1-5 years after operation, pain and cold sensation had no statistically difference (P > 0.05). Intermittent claudication distance had increased. Differences in terms of intermittent claudication distance was statistically significant (Z=43.898, P < 0.001). Significant differences in venous oxygen partial pressure and oxygen saturation were detected between preoperation and 1-year posttransplantation (t values: 36.790, 43.964, P values: 0.040, 0.037). Above results suggest that autologous bone marrow stem cell transplantation for thromboangiitis obliterans obtained stable long-term outcomes.   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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Pioglitazone administration combined with bone marrow mesenchymal stem cells transplantation improved the heart function of rats with myocardial infarction  Wu Quan-hua, Hou Jing-ying, Guo Tian-zhu, Zhong Ting-ting, Long Hui-bao, Xing Yue, Zhou Chang-qing, Zheng Shao-xin, Wang Tong 2015, 19 (23):  3698-3704.  doi: 10.3969/j.issn.2095-4344.2015.23.016 Abstract ( 266 )   PDF (5270KB) ( 291 )   Save BACKGROUND: Our previous work has demonstrated that bone marrow mesenchymal stem cells (BMSCs) transplantation can improve the heart function of rats with myocardial infarction. However, the overall efficacy is not satisfactory. OBJECTIVE: To adopt pioglitazone as a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist combined with BMSCs transplantation therapy, thereby further improving cardiac function of rats with myocardial infarction as well as investigating the relevant mechanisms. METHODS: Twenty Sprague-Dawley rats with myocardial infarction were induced by the left anterior descending coronary artery ligation. The animals were randomized into two groups: BMSCs and BMSCs+pioglitazone. Two weeks later, all the animals received the injection of BMSCs labeled with PKH26 in PBS into the local infarct zone, and then pioglitazone (3 mg/kg/d) was given by the oral gavage for 2 weeks in the BMSCs+pioglitazone group after the cell transplantation. After 2 weeks of cell transplantation, cardiac functions were evaluated by echocardiography. The expressions of PPAR-γ, Connexin 43 and molecules in TGF-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart using immunofluorescent staining, western blot assay and qRT-PCR. RESULTS AND CONCLUSION: There were no differences in the baseline parameters of cardiac function between the two groups. At 2 weeks after cell transplantation, the left ventricular internal diameter at end-diastole, left ventricular internal diameter at end-systole and left ventricular ejection fraction were significantly improved in the BMSCs+ pioglitazone group; the expressions of PPAR-γ and Connexin 43 were distinctly increased in different zones of the left ventricle; the levels of TGF-β1, SMAD2 and SMAD3 were obviously attenuated in the infarct zone and border zone. The above-mentioned findings suggest that pioglitazone, a PPAR-γ agonist, can enhance BMSCs potential in improving the heart function after myocardial infarction, and PPAR-γ may elevate the expression of Connexin 43 via the blockade of the TGF-β1/SMAD signaling pathway in the procedure.  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 bone marrow mononuclear cells from polycythemia vera patients into aplastic anemia mice Zhong Shu-ping, Tian Jing, Liu Xiang, Hou Li-jun, He Zhi-guo, Xu Jing-bo, Li Xue-gang, Xie Feng, Pang Wen-zheng, Liang An-qi 2015, 19 (23):  3705-3710.  doi: 10.3969/j.issn.2095-4344.2015.23.017 Abstract ( 346 )   PDF (4482KB) ( 259 )   Save BACKGROUND: As the high proliferation and low apoptosis of the bone marrow in polycythemia vera patients, hematopoietic stem cells transplanted into NOD/SCID mice can differentiate into erythroid cells, but whether hematopoietic stem cells transplantation could improve the hematopoietic function of aplastic anemia mice is not yet reported. OBJECTIVE: To investigate whether transplantation of bone marrow mononuclear cells with JAK2V617F mutation from polycythemia vera patients can influence hematopoietic reconstruction in aplastic anemia mice. METHODS: Severe aplastic anemia mouse models were established by using recombinant human interferon-γ plus busulfan, and then, these mouse models were randomly divided into experimental group (n=10) and control group (n=10). Bone marrow mononuclear cells isolated from polycythemia vera patients with positive JAK2V617F mutation were transplanted into the mice in the experimental group via tail vein at 5 days after drug withdrawal.  The same volume of normal saline was administered to the control group. Routine peripheral blood test, morphology of bone marrow cells, bone marrow biopsy, and percentage of CD45+ cells in the peripheral blood and marrow were determined at 14 days after transplantation. RESULTS AND CONCLUSION: At 14 days after transplantation, pancytopenia occurred in the experimental group, bone marrow smears showed scattered lymphocytes and hematopoietic progenitors, and bone marrow biopsy presented that hematopoietic tissues were reduced and a small amount of granulocyte cells and erythroblasts could be seen, but megakaryocytes were rare. In contrast to the control group, there was no improvement in the hematopoietic function of mice in the experimental group. CD45+ cells were detectable in the peripheral blood and bone marrow in the experimental group, but not in the control group; and a higher percentage of CD45+ cells was measured in the bone marrow than in the peripheral blood of experimental group mice. Experimental findings indicate that bone marrow mononuclear cells from polycythemia vera patients with positive JAK2V617F mutation can be engrafted into aplastic anemia mice, but cannot improve the hematopoietic function of mice. References | Related Articles | Metrics

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The homing of bone marrow mesenchymal stem cells transplanted via the trachea into rats exposed to silica dust  Huang Ming, Zhou Yong-mei, Li Bin, Wu Qi-feng, Zhu Yu-feng, Liang Wei-hui 2015, 19 (23):  3711-3715.  doi: 10.3969/j.issn.2095-4344.2015.23.018 Abstract ( 171 )   PDF (5502KB) ( 231 )   Save BACKGROUND: Bone marrow mesenchymal stem cells transplantation via the trachea can relieve the lung injury of rats exposed to silica dust, but their distribution and migration in vivo is still unclear. OBJECTIVE: To investigate the distribution and homing of bone marrow mesenchymal stem cells transplanted via the trachea into rats exposed to silica dust. METHODS: Bone marrow mesenchymal stem cells from Sprague-Dawley rats were isolated through bone marrow adherent method and transfected with lentivirus carrying enhanced green fluorescent protein gene (Lv-eGFP). Trypan blue staining and cell counting kit-8 were applied to assay the viability and proliferation of the transfected and untransfected cells. Sprague-Dawley rats, SPF level, were randomized into control group and silica dust exposure group. Rats in the two groups were respectively injected via the trachea with 1 mL of sterile  silica dust suspensions (40 g/L) and 1 mL of normal saline. At 2 days after modeling, 2.2×106 transfected bone marrow mesenchymal stem cells were injected via the trachea into the rats of control group and silica dust exposure group. Rats were killed at weeks 1, 2, 3, 4 after transplantation, and the distribution and intensity of green fluorescence in the lung, heart, liver, spleen, kidney, and brain tissue were observed under the fluorescence microscopy by frozen sections and analyzed using imaging analysis software. RESULTS AND CONCLUSION: When the multiplicity of infection was 50, there were no significant differences between the viability and proliferation activity of the transfected and untransfected cells (P > 0.05). After transplantation of transfected bone marrow mesenchymal stem cells, strong green fluorescence was observed widely in the lung, especially around the bronchus and blood vessels, and still obvious at the 4th week. The fluorescence of other organs also could be observed at the 1st week. It was strong and wide in the liver, spleen and heart, while faint and less in the kidney and brain, and all reduced with time. It shows bone marrow mesenchymal stem cells transplanted via the trachea into rats exposed to silica dust can be homing to the injured lung 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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Effect of enamel matrix derivatives on the differentiation and proliferation of human periodontal ligament stem cells  Wang Shuang, Feng Pei-xun, Chen Yue, Zhang Hai-juan, Li Sha, Bao Qing-hong, Guan Li-min 2015, 19 (23):  3716-3722.  doi: 10.3969/j.issn.2095-4344.2015.23.019 Abstract ( 380 )   PDF (5038KB) ( 602 )   Save BACKGROUND: The enamel matrix derivative has been used in the clinical treatment of severe periodontitis; however, the mechanism(s) by which enamel matrix derivative promotes periodontal regeneration is still obscure. OBJECTIVE: To explore the effects of enamel matrix derivatives on the differentiation and proliferation of periodontal ligament stem cells. METHODS: Periodontal ligament stem cells were isolated and identified from human teeth. Cloning forming efficiency, surface antigen expression and pluripotency were detected and identified. Enamel matrix derivatives with different concentrations (20, 50, 100 mg/L) were used to culture periodontal ligament stem cells for 2 and 4 weeks. Collagen synthesis and mineralized nodule formation were detected using Trichrom staining and Von Kosa’s staining, respectively; real-time RT-PCR was employed to detect expressions of collagen type I, osteocalcin, and RUX2; MTT and cell growth rate assay were used to detect the proliferation of periodontal ligament stem cells. RESULTS AND CONCLUSION: Periodontal ligament stem cells were spindle-shaped and showed a higher  colony forming efficiency than periodontal ligament cells. The expressions of surface antigens of periodontal ligament stem cells-CD105, CD29, CD45, CD44 were respectively 99.8%, 99.7%, 1.26%, 98.8%, indicating periodontal ligament stem cells have the multilineage differentiation potential. Enamel matrix derivatives improve the collagen synthesis and mineralization nodule formation of periodontal ligament stem cells in a time-dose dependent manner. They also can improve the expression of osteogensis-related genes collagen type I, osteocalcin, RUX2 and proliferation of periodontal ligament 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 platelet lysate on the biological characteristics of human mesenchymal stem cells   Wu Di, Wu Xiao-yun, Wu Yan 2015, 19 (23):  3723-3728.  doi: 10.3969/j.issn.2095-4344.2015.23.020 Abstract ( 545 )   PDF (5253KB) ( 467 )   Save BACKGROUND: Platelet lysate has been known as a kind of lysate of autologous or allogeneic platelet-rich products. It not only removes the residual cell structure, reduces immunogenicity, but also retains many growth factors. Platelet lysate has been suggested as a substitute for fetal bovine serum to expand mesenchymal stem cells in vitro. OBJECTIVE: To observe the effects of platelet lysate on biological characteristics of human bone marrow mesenchymal stem cells and adipose mesenchymal stem cells, and provide some experimental data for clinical cell therapy and regenerative medicine. METHODS: Platelet lysate was prepared by repeated freezing and thawing from fresh blood. Healthy adult bone marrow and adipose tissue were collected. Human bone marrow mesenchymal stem cells and adipose  mesenchymal stem cells were obtained by density gradient centrifugation and type I collagenase digestion. We tested the morphology, cell phenotype, differentiation characteristics, proliferation capacity, colony forming ability and the level of cytokine secretion of bone marrow mesenchymal stem cells and adipose mesenchymal stem cells after cultured with platelet lysis. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells and adipose mesenchymal stem cells were successfully cultured in vitro using platelet lysate. There were no significant differences in morphology, cell phenotype, colony forming ability and the level of cytokine secretion, and chondrogenic, osteogenic and adipogenic capacities between bone marrow mesenchymal stem cells and adipose mesenchymal stem cells. Adipose mesenchymal stem cells had a high cumulative population doublings than bone marrow mesenchymal stem cells (P < 0.05). These findings suggest adipose mesenchymal stem cells had a stronger proliferative ability, and are more suitable for large-scale expansion in vitro cultivation system of platelet lysate compared with 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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Effects of umbilical cord Wharton’s jelly mesenchymal stem cell transplantation on the expression of inflammatory factors in rats with spinal cord injury  Ma Shan-shan, Qu Rui-na, Tian Yi, Yao Ning, Cui Yuan-bo, Han Kang, Xing Qu, Yang Bo, Guan Fang-xia 2015, 19 (23):  3729-3735.  doi: 10.3969/j.issn.2095-4344.2015.23.021 Abstract ( 186 )   PDF (5543KB) ( 327 )   Save BACKGROUND: The production and release of a large amount of inflammatory factors caused by immune system inflammatory response mainly contributes to secondary spinal cord injury. OBJECTIVE: To investigate the effects of umbilical cord Wharton’s jelly mesenchymal stem cell transplantation on repair of injured neurological function and expression of inflammatory factors monocyte chemoattractant protein 1 and interleukin 10 in rats with acute spinal cord injury. METHODS: Eighty-one healthy adult male Sprague-Dawley rats were randomly and equally divided into sham operation, model and cell transplantation groups, with 27 rats per group. Rats in the latter two groups were subjected to hemisection of the spinal cord to establish acute spinal cord injury models. Rat models in the cell  transplantation group received umbilical cord Wharton’s jelly mesenchymal stem cell injection (1×106) via the tail vein. Rat neurological function was evaluated using the BBB score at different time points after spinal cord injury. The expression of monocyte chemoattractant protein 1 and interleukin 10 in injured spinal cord tissue was detected using ELISA assay at different time points after spinal cord injury. Migration and neuronal differentiation of umbilical cord Wharton’s jelly mesenchymal stem cells in the injured spinal cord tissue were determined using immunohistochemical staining method. RESULTS AND CONCLUSION: Compared with the sham operation and model groups, rat neurological function was significantly recovered in the cell transplantation group (P < 0.05). Compared to the model group, monocyte chemoattractant protein 1 level in the serum and monocyte chemoattractant protein 1 mRNA and protein expression in the injured spinal cord tissue were significantly lower (P < 0.05), but interleukin 10 mRNA and protein expression in the injured spinal cord tissue was significantly higher (P < 0.05), in the cell transplantation group. In the cell transplantation group, umbilical cord Wharton’s jelly mesenchymal stem cells could migrate to the injured region and express glial fibrillary acidic protein. These findings suggest that umbilical cord Wharton’s jelly mesenchymal stem cells promote rat neurological function recovery by regulating the inflammatory response in the injured spinal cord tissue, which is likely to be one of mechanisms by which transplantation of umbilical cord Wharton’s jelly mesenchymal stem cells treats 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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Biocompatibility of porcine bone marrow mesenchymal stem cells-derived bile duct endothelial cells with electrospun nanofibers Yang Yang, Zhou Jia-hua, Yin Xue-yan, Xu Yong, Cao Yang2, Xu Qian 2015, 19 (23):  3736-3743.  doi: 10.3969/j.issn.2095-4344.2015.23.022 Abstract ( 320 )   PDF (2425KB) ( 274 )   Save BACKGROUND: Repair of extrahepatic biliary tract injury is a difficult problem in the abdominal surgery. Tissue-engineered extrahepatic biliary tract is an ideal selection for this problem. Construction of tissue-engineered extrahepatic biliary tract with excellent performance is a key to related studies. OBJECTIVE: To investigate the biocompatibility of bile duct endothelial cells differentiated by porcine bone marrow mesenchymal stem cells with electrospun nanofibers. METHODS: Porcine bone marrow mesenchymal stem cells were induced toward biliary tract endothelial cells, which were then identified by morphology and RT-PCR. Polylactic-co-glycolic acid (PLGA) nanofiber membranes were prepared by electrospinning. The morphology was determined by scanning electron microscopy and the short-term (2-week) in vitro degradation rate was determined. Adhesion and proliferation of biliary tract endothelial cells on the nanofiber surface was analyzed by calculating the cell adhesion rate and MTT assay, respectively. Cell growth, morphology and distribution on the material surface were observed by fluorescence staining and scanning electron microscopy, respectively. RESULTS AND CONCLUSION: After 4 weeks of directed differentiation of bone marrow mesenchymal stem cells in vitro, cells showed typical morphology of dendritic bile duct endothelial cells and had the expression of CK19. Scanning electron micrographs showed that electrospun materials were continuous nanofibers with diameters between 200 and 500 nm. No significant degradation of the PLGA nanofibers was observed within 2 weeks. Based on the measured cell adhesion rate, MTT assay, fluorescence staining, and scanning electron microscopy, the differentiated cells possessed a good proliferative capacity on PLGA nanofibers. Bone marrow mesenchymal stem cells differentiated into bile duct endothelial cells in vitro. Materials prepared by the electrospinning method had a nanofiber structure, which did not significantly degrade within 2 weeks. Differentiated cells exhibit good biocompatibility with the nanofibers. 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 magnetic fields on the differentiation of bone marrow mesenchymal stem cells Zhang Jing, Zhan Qi, Huang Yan 2015, 19 (23):  3744-3749.  doi: 10.3969/j.issn.2095-4344.2015.23.023 Abstract ( 274 )   PDF (739KB) ( 399 )   Save null Figures and Tables | References | Related Articles | Metrics

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Hypoxia inducing factor 1 alpha-transfected cardiac stem cells in repair of necrotic myocardium  Li Sha, Li Shu-ren, Zhang Qian-hui 2015, 19 (23):  3750-3754.  doi: 10.3969/j.issn.2095-4344.2015.23.024 Abstract ( 261 )   PDF (917KB) ( 427 )   Save BACKGROUND: Cardiac stem cells transplanted to the myocardial infarction area can effectively improve ventricular remodeling and promote heart function. But the survival rate of transplanted cells is lower in the infracted area under hypoxic microenvironment. Hypoxia inducing factor 1 alpha under anoxic conditions can stably express, and meanwhile increase the activity and survival ability of myocardial cells. OBJECTIVE: To elaborate the research progress in hypoxia inducing factor 1 alpha-transfected cardiac stem cells for treatment of myocardial infarction from the following aspects: cardiac stem cell characteristics, mechanism underlying myocardial protection of hypoxia inducing factor 1 alpha, selection of carriers and transplantation approach.  METHODS: A computer-based search of CNKI and PubMed was performed for articles related to cardiac stem cells and hypoxia inducing factor 1 alpha published from January 2000 to January 2015. The keywords were “cardiac stem cells, hypoxia inducible factor 1(HIF-1a), gene delivery” in Chinese and English, respectively, which appeared in the title and abstract. Finally, 37 relevant articles were enrolled in result analysis. RESULTS AND CONCLUSION: Several studies have confirmed that hypoxia inducing factor 1 alpha can improve the survival rate of cardiac stem cells under anoxic conditions. Increasing evidences from animal experiments have shown that cardiac stem cells and hypoxia inducing factor 1 alpha exert protective and repairing effects on myocardial infarction. Currently, there is no successful report about hypoxia inducing factor 1 alpha gene transfection of cardiac stem cells, but relevant studies are proceeding. Gene modified cardiac stem cells are expected to 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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Umbilical cord blood stem cell transplantation and endovascular treatment of diabetic lower limb ischemic disease  Ma Hong-fang, Wang Fu-jun 2015, 19 (23):  3755-3760.  doi: 10.3969/j.issn.2095-4344.2015.23.025 Abstract ( 310 )   PDF (918KB) ( 334 )   Save BACKGROUND: In recent years, interventional therapy because of its high success rate, good curative effect, little trauma has been widely recognized, but vascular intimal injury due to interventional therapy still has no effective treatment, resulting in long-term postoperative severe restenosis. Stem cell transplantation has brought a new drawn for the repair of impaired endothelial function, reendothelialization and aided restoration of the blood supply to ischemic tissues. OBJECTIVE: To summarize the recent progress in umbilical cord blood stem cells transplantation and endovascular treatment for treatment of diabetic lower limb ischemic disease. METHODS: A computer-based search of PubMed and CNKI was performed for relevant articles published from January 2000 to March 2015 using the keywords of “umbilical cord blood stem cell transplantation, percutaneous transluminal angioplasty, diabetic lower limb ischemia, restenosis” in English and Chinese. Totally 216 relevant literatures were initially obtained, and 44 articles were ultimately included. RESULTS AND CONCLUSION: By comparing the therapeutic efficacy and long-term prognosis of umbilical cord blood stem cell transplantation and endovascular treatment, it is concluded that umbilical cord blood stem cell transplantation combined with endovascular treatment is better than either of them alone, which can promote the establishment of collateral circulation, improve the impaired endothelial function and reduce the occurrence of restenosis after interventional therapy. 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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Induced pluripotent stem cells in bone tissue regeneration: how to achieve clinical application in orthopedic surgery as soon as possible?  He Rui-xuan, Zhao Liang 2015, 19 (23):  3761-3767.  doi: 10.3969/j.issn.2095-4344.2015.23.026 Abstract ( 324 )   PDF (960KB) ( 368 )   Save BACKGROUND: Biomaterial combined with stem cells is revealing a bright future in bone tissue regeneration in orthopedic surgery. Induced pluripotent stem cells have better cell sources and characteristic, which have become a hotspot of stem cell field. OBJECTIVE: To review the research history, preparation method, cell characteristics of induced pluripotent stem cells and the research developments in orthopeadic surgery so far. METHODS: A computer-based search of CNKI and PubMed was performed for articles about induced pluripotent stem cells and their applications in the field of orthopeadic surgery published from 1999 to 2014. Typical and creative research achievements were enrolled in result analysis. RESULTS AND CONCLUSION: Studies have shown that induced pluripotent stem cells have a potential application prospect in bone tissue regeneration, and they also show a satisfying biocompatibility with various scaffold materials, in which, the induced pluripotent stem cells can maintain a good osteogenetic potential. Detection methods and small-molecule compounds have been discovered gradually for cartilage regeneration. But how to effectively induce the chondrogenic differentiation of induced pluripotent stem cells and to realize the clinical applications need further research. 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 | Related Articles | Metrics

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Fate of reprogramming cells Ruan Guang-ping, Yao Xiang, Liu Ju-fen, Hu Yuan-yuan, Wang Jin-xiang, He Jie, Zhao Jing, Pan Xing-hua 2015, 19 (23):  3768-3772.  doi: 10.3969/j.issn.2095-4344.2015.23.027 Abstract ( 191 )   PDF (949KB) ( 248 )   Save BACKGROUND: Somatic cell reprogramming technology, also known as recombinant technology, has completed differentiated somatic cells back to the original totipotent or pluripotent state, and can be re-differentiated into cells different from original ones. Re-programming techniques are able to harvest specifically induced pluripotent stem cells and disease-specific induced pluripotent stem cells from patients, which can significantly reduce the immune rejection. OBJECTIVE: To explore the method from direct reprogramming to specific cell lines and to conclude the molecular mechanisms underlying reprogramming. METHODS: A computer-based search of VIP, Wanfang, CNKI, PubMed and Springer databases was performed for articles related to cell reprogramming techniques published from January 1958 to April 2015 using the keywords of “reprogramming” in Chinese and English, respectively. After elimination of unrelated and repetitive studies, 40 articles were retained for further analysis. RESULTS AND CONCLUSION: Current reprogramming steps are inefficient that in the specific group only a relatively small number of cells can be reprogrammed, and the extent and integrity of reprogramming are questionable. Direct reprogramming of adult cells and specific cell lines that are changed to another line is always a difficulty in developmental biology. Recent studies have proved that differentiated cells can forcibly express specific transcription factor to improve cell differentiation. This discovery is a great progress in regenerative medicine that cell replacement therapy can be provided for renewable disorders. At present, the basic molecular mechanisms require further clarification, and there are many problems to be solved before the direct reprogramming is used clinically. 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 | Related Articles | Metrics