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05 March 2015, Volume 19 Issue 10 Previous Issue    Next Issue

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Improved primary culture system increases the yield of human umbilical cord mesenchymal stem cells Gao Yan-lin, Zhang Ning-kun, Chen Hou-liang, Gao Lian-ru, Zhu Zhi-ming 2015, 19 (10):  1477-1481.  doi: 10.3969/j.issn.2095-4344.2015.10.001 Abstract ( 422 )   PDF (453KB) ( 614 )   Save BACKGROUND: At present, a lot of research about culture methods for umbilical cord mesenchymal stem cells, but not for the waste of primary system. OBJECTIVE: To explore the best culture method of human umbilical cord mesenchymal stem cells in vitro. METHODS: Human umbilical cord mesenchymal stem cells were prepared by tissue explants method, recorded as initial culture group. The centrifugal fluid and tissue of the primary culture flask were centrifuged and divided into three groups for secondary culture: tissue group, mixed group and pure liquid group. Cell morphology, time for cell acquisition, and yield of primary cells in the four groups were observed; the cell growth curve was analyzed by MTT assay; and cell cycle and phenotype were detected by flow cytometry. RESULTS AND CONCLUSION: The average time for cell acquisition in the initial culture group, tissue group, mixed group and pure liquid group were (15.00±0.45), (7.0±0.3), (8.00±0.25) and (8.00±0.25) days, respectively. The number of cells at first generation was (4.0±0.5)×105, (9.0±0.55)×105, (15.0±0.2)×105 and (7.0±0.33)×105 per T75 culture bottle, respectively. Under the inverted microscope, cells in the four groups were fusiform-like adherent cells, which were in parallel or circinate arrangement. Growth curve, proliferative activity, surface markers of the four groups had no significant differences. The human umbilical cord mesenchymal stem cells can be obtained rapidly and largely through the secondary culture to the primary culture system. 1库： 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 全文链接： Figures and Tables | Related Articles | Metrics

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5-Fluorouracil enriches cancer stem cells in tongue squamous cell carcinoma Tca8113 Xu Wang, Ji Ping 2015, 19 (10):  1482-1487.  doi: 10.3969/j.issn.2095-4344.2015.10.002 Abstract ( 304 )   PDF (496KB) ( 434 )   Save BACKGROUND: Tumor cells are resistant to chemotherapeutic drugs, and drug resistance is closely correlated with tumor stem cells. Therefore, how to kill tumor stem cells will become the key to the treatment of oral squamous cell carcinoma. OBJECTIVE: To study the effect of 5-fluorouracil on biological characteristics of tongue squamous cell carcinoma Tca8113 cells. METHODS: Viability of Tca8113 cells treated with different concentrations of 5-fluorouracil was determined by cell counting kit-8, and the best drug concentration and time were screened for subsequent experiments. Tca8113 cells without 5-fluorouracil acted as control group. Then the cell cycle and percentage of the side population cells in Tca8113 cells were determined by flow cytometry. Scratch test was used to determine the migration ability of Tca8113 cells. RESULTS AND CONCLUSION: Results from cell counting kit-8 showed that 5-fluorouracil inhibited the viability of Tca8113 cells positively in a time- and dose-dependent manner. Tca8113 cells under intervention with 50 mg/L 5-fluorouracil for 48 hours showed lowest cell viability. Flow cytometry results showed that in the experimental group, G0/G1 phase cells increased significantly compared with the control group (P=0.01), S phase cells decreased significantly compared with the control group (P=0.244), and G2/M phase cells disappeared completely. After treatment with 5-fluorouracil, the percentage of side population cells was increased significantly (P=0.00). The scratch test showed that in the experimental group, the cells had better ability of wound healing than those in the control group. In conclusion, 5-fluorouracil can enrich the cancer stem cell population in Tca8113 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Lentivirus-mediated cyclooxygenase 2 and aggrecanase 1 silencing and insulin-like growth factor 1 overexpression in human bone marrow mesenchymal stem cells Yuan Yu-qi, Zhang Hai-ning, Kong Xia, Sui Ai-hua, Wang Ying-zhen 2015, 19 (10):  1488-1494.  doi: 10.3969/j.issn.2095-4344.2015.10.003 Abstract ( 402 )   PDF (392KB) ( 451 )   Save BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage.   OBJECTIVE: To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cells. METHODS: Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cells cultured in vitro (experimental group). The human bone marrow mesenchymal stem cells transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cells transfected with no treatment served as blank group. RESULTS AND CONCLUSION: Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cells were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfully silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cells, which brings a new hope for the systemic gene treatment of arthritis. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Functional status of bone marrow mesenchymal stem cells from osteoarthritis patients and disease development Wang Zhi-zhou, Xu Chao, Han Ya-jun, Tie Xiao-jia, Guo Hong-liang, Yilihamu Tuoheti, Wang Jun-feng, Yan Bin, Bi Xiao-juan, Guan Zhen-peng 2015, 19 (10):  1495-1500.  doi: 10.3969/j.issn.2095-4344.2015.10.004 Abstract ( 292 )   PDF (1943KB) ( 401 )   Save BACKGROUND: Bone marrow mesenchymal stem cells are crucial for bone and cartilage development and regeneration at a cellular level. Insufficient quantity and functional impairment of bone marrow mesenchymal stem cells is widely considered to be one of osteoarthritis causes. OBJECTIVE: To explore the relationship between the functional status of bone marrow mesenchymal stem cells and disease progression in osteoarthritis patients. METHODS: Thirty patients with osteoarthritis were enrolled from July 2013 to October 2014, and divided into control, mild osteoarthritis, and severe osteoarthritis groups, with 10 cases in each group. 5 mL bone marrow from the femur or tibia was extracted from each patient to isolate and culture bone marrow mesenchymal stem cells. Proliferation ability of cells at passage 3 was detected using cell counting kit-8; toluidine blue staining was performed at 14 days after chondrogenic induction; real-time PCR was used to detect the mRNA expression of Aggrecan and Col2A1 in the control group after chondrogenic induction. RESULTS AND CONCLUSION: After in vitro culture, bone marrow mesenchymal stem cells grew adherently in polygonal and fusiform shape with multiple processes at uniform size. The cytoplasm contained larger particles and the nuclei were ovoid. Most of cells were in cell division phase. The proliferation ability was strongest in the control group and weakest in the severe osteoarthritis group. Cells from the three groups were all at plateau phase after 1 week culture. At 14 days after chondrogenic induction, the cells were polygonal and quasi-circular, and purple metachromatic granules distributed outside of the cytoplasm. The expression of Aggrecan and Col2A1 in the control group displayed an overexpression trend. These findings indicate that the functional status of bone marrow mesenchymal stem cells from osteoarthritis patients is negatively correlated with the severity of disease, which can influence the disease progression in osteoarthritis patients. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Osteogenic induction of human bone marrow mesenchymal stem cells cultured in complex medium Xu Li-li, Sun Xiao-juan, Hao Xiu-xian, Xie Ting-ting, Yang Nai-long 2015, 19 (10):  1501-1501.  doi: 10.3969/j.issn.2095-4344.2015.10.005 Abstract ( 385 )   PDF (1742KB) ( 361 )   Save BACKGROUND: Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis.   OBJECTIVE: To observe the osteogenic differentiation of human bone marrow mesenchymal stem cells cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS: Mesenchymal stem cells were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cell surface markers was detected by flow cytometry. Cell ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cells were cultured in osteogenic induction medium containing dexamethasone, vitamin C and β-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND CONCLUSION: A large number of adherent cells were visible as fibrous growth at 2 weeks after culture and strongly expressed CD44, CD29, but did not express CD34, CD45. These cells could be induced to differentiate into osteoblasts, and express bone morphogenetic protein-2 mRNA. Alizarin red staining and alkaline phosphatase staining were positive for the cells. These findings suggest that human bone marrow mesenchymal stem cells cultured in dexamethasone, vitamin C and beta-glycerophosphate can differentiate into osteoblasts, and has a potential for the treatment of osteoporosis. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Chondrogenic differentiation of mesenchymal stem cells on matrigel/chitosan scaffolds Dai Yun, He Jing, Wu Fang 2015, 19 (10):  1506-1510.  doi: 10.3969/j.issn.2095-4344.2015.10.006 Abstract ( 374 )   PDF (2556KB) ( 559 )   Save BACKGROUND: Previous studies have found that the basement membrane matrix can induced the chondrogenic differentiation of bone marrow mesenchymal stem cells, but there is a wide gap between its mechanical properties and practical application, and further research is needed. OBJECTIVE: To prepare a suitable matrigel/chitosan scaffold that has appropriate mechanical properties and remarkable bioactivity for cartilage repair.  METHODS: We selected genipin as cross-linking agent, and mixed Matrigel with cross-linked chitosan at different ratios (2:1, 1:1, 1:3). Then rat bone marrow mesenchymal stem cells were seeded on different scaffolds and cultured for 14 days. The mechanical properties of materials were measured by DMA. Cell counting kit-8, FDA staining, ELISA kits and Alcian blue staining were used to measure the bioactivity of materials. RESULTS AND CONCLUSION: The storage modulus of scaffolds was raised from 0.48 kPa to 1.78 kPa with increase ratio of chitosan. Cells spread well in the early period on all scaffolds, and then the cells on the chitosan scaffold showed reduced chondrogenic activity, but cells on the scaffolds with matrigel could maintain chondrogenic differentiation. The matrigel/chitosan scaffold at a ratio of 1:1 had appropriate mechanical properties and higher levels of collagen II and collagen X at 14 days. The prepared matrigel/chitosan scaffold with decent mechanical performance can promote the differentiation of bone marrow mesenchymal stem cells into chondrogenic lineages, which can be used in cartilage tissue engineering. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Biological effect of human bone marrow mesenchymal stem cells on hepatocellular carcinoma cells Chen Qi-feng, Fang Xiao-ming, Yao Ning, Fang Xu-dong, Gong Mou-chun 2015, 19 (10):  1511-1515.  doi: 10.3969/j.issn.2095-4344.2015.10.007 Abstract ( 288 )   PDF (1846KB) ( 326 )   Save  BACKGROUND: The metastatic potential of hepatocellular carcinoma cells is key factor influencing patient’s prognosis. To observe the effect of human bone marrow mesenchymal stem cells on metastasis of hepatocellular carcinoma is of great significance for improving the lifetime of hepatocellular carcinoma patients. OBJECTIVE: To explore the biological effect of human bone marrow mesenchymal stem cells on hepatocellular carcinoma cells with different metastatic potentials. METHODS: Human bone marrow mesenchymal stem cells and hepatocellular carcinoma cell suspension with high and low metastatic potentials were respectively injected into the Transwell chamber, and after 36 hours of co-culture, ELISA method was used to detect the absorbance value as well as cell counting method was used to observe the changes in the invasion ability of hepatocellular carcinoma cells. The effects of human bone marrow mesenchymal stem cells on the proliferation of hepatocellular carcinoma cell suspension with high and low metastatic potentials were determined using cell counting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration (alpha V), transforming growth factor beta 1 and programmed cell death protein 5. RESULTS AND CONCLUSION: (1) The number of migrated hepatocellular carcinoma cells was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group (P < 0.05). (2) The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential (P < 0.05), but there was no change in the expression of integration (alpha V) (P > 0.05). In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration (alpha V) were declined remarkably (P < 0.05). (3) Results from the semi-quantitative detection of proliferation ability showed that the absorbance value of the co-culture group was significantly higher than that of the single culture group(P < 0.05). (4) In the co-culture group with high metastatic potential, the expression of transforming growth factor beta 1 was up-regulated significantly (P < 0.05), but the expression of programmed cell death protein 5 showed no changes(P > 0.05). However, in the co-culture group with low metastatic potential, the expression of transforming growth factor beta 1 and programmed cell death protein 5 was both increased dramatically (P < 0.05). These findings suggest that the human bone marrow mesenchymal stem cells reduce the invasion ability of hepatocellular carcinoma cells, and enhance their ability of proliferation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Long-term effects of cardiac stem cells transplantation to improve the electrophysiological stability and ventricular fibrillation threshold after myocardial infarction Chen Li-peng, Yan Ping, Guo Tian-zhu, Hou Jing-ying, Zheng Shao-xin, Zhou Chang-qing,Long Hui-bao, Zhong Ting-ting, Wang Tong 2015, 19 (10):  1516-1522.  doi: 10.3969/j.issn.2095-4344.2015.10.008 Abstract ( 494 )   PDF (2456KB) ( 466 )   Save BACKGROUND: Our previous work has demonstrated that the electrophysiological stability and ventricular fibrillation threshold were distinctly improved in rats with myocardial infarction 6 weeks after cardiac stem cells transplantation (short- to mid-term).   OBJECTIVE: To observe the long-term effects of cardiac stem cells transplantation on the ventricular fibrillation threshold and electrophysiological stability in rats with heart failure post myocardial infarction. METHODS: Twenty Sprague-Dawley rats with myocardial infarction were induced by the left anterior descending coronary artery ligation. Two weeks later, animals were randomized to receive the injection of cardiac stem cells labeled with PKH26 in PBS or PBS alone into the local infarct zone. Twelve weeks after the cardiac stem cells or PBS injection, the electrophysiological characteristics and ventricular fibrillation threshold were measured at the infarct zone, the infarct marginal zone and the non-infarct zone. Labeled cardiac stem cells were detected and the expression of connexin-43 and α-sarcomeric actin was examined in cryostat sections from each harvested heart. RESULTS AND CONCLUSION: Compared with the PBS group, no significant differences were revealed in the unipolar electrogram activation recovery time and the correct activation recovery time in the infarct zone, the infarct marginal zone and the non-infarct zone in the cardiac stem cells group at 12 weeks post the cell transplantation (P > 0.05). There were no remarkable differences in the correct activation recovery time dispersion and induced malignant ventricular arrhythmias between the two groups (P > 0.05). No obvious differences were discovered in the ventricular fibrillation threshold in the infarct zone and infarct marginal zone in the cardiac stem cells group in contrast to the PBS group (P > 0.05), though a statistical significance could be found in the non-infarct zone between the two groups (P < 0.05). Labeled cardiac stem cells were identified in the infarct marginal zone and expressed connexin-43 and α-sarcomeric actin. It suggested that the beneficial effects of cardiac stem cells in modulating the electrophysiological stability and improving the ventricular fibrillation threshold faded away 12 weeks after cell transplantation. However, the mechanisms involved remained unclear. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Human lung adenocarcinoma stem cell phenotypes and patient’s prognosis Maierdan Sailimu, Alidan Erkin, Jin Zhong, He Tie-han 2015, 19 (10):  1523-1527.  doi: 10.3969/j.issn.2095-4344.2015.10.009 Abstract ( 354 )   PDF (390KB) ( 519 )   Save BACKGROUND: Lung adenocarcinoma stem cells are a very important marker for diagnosis of lung adenocarcinoma. OBJECTIVE: To investigate the relationship between human lung adenocarcinoma stem cell phenotypes and patient’s prognosis. METHODS: From February 2010 to January 2013, 48 patients with lung adenocarcinoma admitted in the First Affiliated Hospital of Xinjiang Medical University were enrolled. Lung adenocarcinoma cancer stem cell phenotypes were detected with immunofluorescence method, and the relationship of different phenotypes and clinical characteristics with patient’s prognosis was compared. RESULTS AND CONCLUSION: SP-C and CCSP expression was observed in the lung adenocarcinoma tissues of 48 cases, possessing the phenotypic characteristics of bronchioloalveolar stem cells. Of the 48 patients, OCT4 was positive in 34 cases (OCT4+ group) and negative in 14 cases (OCT4- group). There was no difference between the two groups in terms of patient’s age, gender, smoking history and tumor stage (P > 0.05). There were 23 cases (68%) in the OCT4+ group with cancer cell metastasis, which was significantly higher than that in OCT4- group (21%; P > 0.05). In the aspects of patient’s age, gender, smoking history, staging and cancer metastasis and other clinical pathological stratification, the 2-year survival rate in the OCT4- group were generally higher than that in the OCT4+ group, and there was a significant difference in the survival curves of two groups (P < 0.05). Human lung adenocarcinoma stem cells have the phenotypic characteristics of bronchioloalveolar stem cells, and meanwhile, if the cells are positive for OCT4, patient’s prognosis is poor and associated with lung adenocarcinoma metastasis. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Intravenous and intramyocardial administration of bone marrow mesenchymal stromal cells cannot improve heart function in a rat model of chronic myocardial infarction Wang Wei, Liu De-bin, Feng Ying, Zhang Hao, Wang Shi-xiong, Gao Bing-ren, Zhang Jing,Wang Wei-fan 2015, 19 (10):  1528-1532.  doi: 10.3969/j.issn.2095-4344.2015.10.010 Abstract ( 308 )   PDF (841KB) ( 396 )   Save BACKGROUND: It has been demonstrated to be effective for the improvement of heart function after acute myocardial infarction with intravenous or intramyocardial administration of bone marrow mesenchymal stromal cells. However, little is known regarding the effect of the combination of intravenous and intramyocardial administration of mesenchymal stromal cells on the heart function of a chronic myocardial infarction model.   OBJECTIVE: To study the effect of intravenous and intramyocardial administration of bone marrow mesenchymal stromal cells on the heart function of a rat chronic myocardial infarction model and the relevant mechanism. METHODS: Bone marrow mesenchymal stromal cells isolated from Lewis rats were expanded ex vivo. BrdU-labeled bone marrow mesenchymal stromal cells (3×106) were administered via the femoral vein and the myocardial surface respectively into rat models of chronic myocardial infarction in cell transplantation group. The equal volume of PBS was injected into the same place in control group. Four weeks after injection, echocardiography was performed to evaluate the heart function, and then the heart tissues were harvested for immunohistochemistry examination. The total blood vessel density in the scar area was evaluated. RESULTS AND CONCLUSION: At 4 weeks after cell implantation, the left ventricular function was not improved in the two groups. The immunohistochemistry staining showed that (1) the mesenchymal stromal cells in the myocardium did not differentiate to myocardial cells; (2) there was no significant difference in the total blood vessel density in the scar area between the cell transplantation and control groups. Taken together, the combined intravenous and intramyocardial administration of bone marrow mesenchymal stromal cells cannot improve heart function in a rat chronic myocardial infarction model. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Umbilical cord mesenchymal stem cells with bone marrow stem cells in the treatment of decompensated cirrhosis: a 1-year follow-up study Zeng Zhi-yu, Li Dong-liang, Fang Jian, Ruan Mei, Xia Lei, Zhang Zhi-qiang, Zhang Shi-an 2015, 19 (10):  1533-1538.  doi: 10.3969/j.issn.2095-4344.2015.10.011 Abstract ( 317 )   PDF (369KB) ( 509 )   Save BACKGROUND: There are most single-center studies about bone marrow stem cells applied to treat decompensated cirrhosis, but the therapeutic results are not ideal. It is possibly related to aging, physical weakness, poor bone marrow hematopoietic function, less available number of stem cells and feeble ability of regeneration and proliferation in liver cirrhosis patients. Umbilical cord mesenchymal stem cells are characterized of easy to obtain, wide source and weak immunogenicity. Co-transplantation of bone marrow stem cells and umbilical cord mesenchymal stem cells may improve the therapeutic effects on decompensated cirrhosis patients. OBJECTIVE: To investigate the efficacy and safety of co-transplantation of umbilical cord mesenchymal stem cells and bone marrow stem cells on decompensated cirrhosis. METHODS: Thirty-two decompensated cirrhosis patients were randomly divided into two groups: in stem cell group, 13 patients received co-transplantation of umbilical cord mesenchymal stem cells and bone marrow stem cells based on regular medical treatment; in control group, 19 patients only underwent the regular medical treatment. All the patients were follow-up for 1 year. Alanine aminotransferase, albumin, total bilirubin, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score, 1-year survival rate, Quality of Life score and adverse reactions related to stem cell therapy were observed and recorded in the two groups at 4, 12, 52 weeks after treatment. RESULTS AND CONCLUSION: At 4, 12, 52 weeks after treatment, improvements in the liver function, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score were found in the two groups, but there was no difference between the two groups (P > 0.05). At 4 weeks after transplantation, the clinical symptoms and Quality of Life score in the stem cell group were significantly improved, which were better than those in the control group (P < 0.05). But at 12 and 52 weeks after treatment, no difference was found between the two groups (P > 0.05). In addition, the 1-year survival rate showed no difference between the two groups, and no severe adverse reactions related to stem cell therapy occurred during the follow-up. Co-transplantation of umbilical cord mesenchymal stem cells and bone marrow stem cells is safe and effective to improve the clinical symptoms of decompensated cirrhosis patients. However, further studies with larger samples are warranted to better clarify the co-transplantation 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Lentiviral vectors with red fluorescent proteins for transfection of human bone marrow mesenchymal stem cells: the red fluorescent protein can express for 21 consecutive days and exert no effects on cell viability Yin Biao, Yang Yi-xi, Yang Bo, Wang Le, Li Shuang-qing, Yang Fu-guo 2015, 19 (10):  1539-1543.  doi: 10.3969/j.issn.2095-4344.2015.10.012 Abstract ( 325 )   PDF (1910KB) ( 339 )   Save  BACKGROUND: To grasp the optimal multiplicity of infection (MOI) and the time when stronger fluorescence intensities produce can lay the foundation for tracing observation of human bone marrow mesenchymal stem cells in vivo in animal models. OBJECTIVE: To investigate the feasibility of HIV-1 lentivirus carrying enhanced red fluorescent protein to transfect human bone marrow mesenchymal stem cells. METHODS: Human bone marrow mesenchymal stem cells at passage 4 were divided into blank group and MOI 2, 3, 4 groups. After that, the cells were seeded into 12-well plates at a density of 5.0×105, and cultured in 1 mL complete medium for adult bone marrow mesenchymal stem cells containing 1% fetal bovine serum. The infectious titer of lentivirus-carried enhanced red fluorescent protein was adjusted to 1.0×1011 TU/L. Lentivirus solution 10, 15, 20 μL at MOI=2, 3, 4 were respectively added into the MOI 2, 3, 4 groups, and 10 μL PBS was added into the blank group. At 24 and 72 hours after transfection, the expression of red fluorescence was observed under an inverted fluorescence microscope and the transfection efficiency was calculated. RESULTS AND CONCLUSION: Enhanced red fluorescent protein expressed stably in bone marrow mesenchymal stem cells. At 24 hours after transfection, red fluorescence could be seen under the inverted fluorescence microscope and achieved the peak at 72 hours after transfection. Within 21 days after transfection, there were no differences in the number of human bone marrow mesenchymal stem cells between the MOI 2, 3, 4 groups and blank group (P > 0.05). These results show that the HIV-1 lentivirus carrying enhanced red fluorescent protein is feasible to transfect human bone marrow mesenchymal stem cells, with the highest transfection efficiency when the MOI=4, which can express at least for 21 days and have no effects on the proliferative activity of labeled 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of bone marrow mesenchymal stem cell conditioned medium with and without inflammatory activation on radiation-induced intestinal injury Liu Wan-wei, Chen Yun, Zheng Yue, Sha Wei-hong, Wang Qi-yi, Ye Shan-liang, Chen Hao 2015, 19 (10):  1544-15550.  doi: 10.3969/j.issn.2095-4344.2015.10.013 Abstract ( 398 )   PDF (3008KB) ( 375 )   Save BACKGROUND: Conditioned medium from mesenchymal stem cells (MSC-CM) may represent a promising alternative to MSCs transplantation. Previous studies have shown that inflammatory activation can strengthen the multiple biological potencies of MSCs; however, normal MSCs with insufficiency of immunocompetence and migration ability are not effective for tissue damage repair. OBJECTIVE: To investigate differential effects of MSC-CM with and without inflammatory activation on radiation-induced intestinal injury. METHODS: MSCs from the bone marrow of SD rats were separated, cultured and identified, and then co-cultured with non-irradiated IEC-6 or irradiated IEC-6 in a transwell system for 24 hours. Then, MSCs with inflammatory activation were cultured alone for another 48 hours. After that, the supernatant was collected as non-activated MSC-CM (MSC-CMNOR) and MSC-CM under radiation-induced inflammatory condition (MSC-CMIR). Rats were exposed to 14 Gy whole abdominal irradiation and randomly divided into four groups: control group, radiation injury group (DMEM/F12), MSC-CMNOR group and MSC-CMIR group. Continuous administration was given via tail vein and intraperitoneal implantation of Alzet microosmotic pumps. Intestinal samples were collected at 1, 3, 7 days after radiation for analysis of short circuit variation, at 3 days after radiation for analysis of intestinal epithelium ultrastructure, and at 1, 3, 5, 7, 14 days after radiation for histological observation of the intestinal epithelium using hematoxylin-eosin staining. Blood samples were collected at 1, 3, 7 days after radiation for analysis of serum xylose levels. In addition, the survival state and survival time of rats were observed and recorded. RESULTS AND CONCLUSION: The short circuit variation responding to electrical field stimulation was significantly reduced at all frequencies, but it was significantly improved in the MSC-CMIR group. Similarly, the intestinal absorption (serum xylose levels) was also significantly impaired by irradiation, but improved by delivery of MSC-CMIR (P < 0.05). At 3 days after MSC-CMIR infusion, the intestinal epithelium exhibited an increase in crypt size and villous length (P < 0.05). Under the electron microscope, a reduction in intestinal microvilli and open tight junctions in irradiated intestinal epithelium was found, and the intestine from rats treated with MSC-CMIR had more obvious tight junctions. In addition, treatment with MSC-CMIR dramatically improved the survival rate and mean survival time of irradiated rats as compared to those treated with DMEM/F12 or MSC-CMNOR (P < 0.05). Taken together, the present study demonstrated that MSC-CMIR, but not non-activated MSC-CM, improves the structural and functional restoration of the small intestine after radiation-induced intestinal 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Homing time window of allogeneic bone marrow stem cells transplanted into Chinese miniature pigs with myocardial infarction Xiang Li-ping, Guan Li-na, Shi Qi, Mu Yu-ming 2015, 19 (10):  1551-1555.  doi: 10.3969/j.issn.2095-4344.2015.10.014 Abstract ( 286 )   PDF (1809KB) ( 318 )   Save BACKGROUND: Due to the different transplantation time after myocardial infarction, the homing ability of bone marrow mesenchymal stem cells to damaged tissues as well as the repairing role will be very different.   OBJECTIVE: To explore the optimal window time for the homing of bone marrow mesenchymal stem cells to the myocardial tissue after myocardial infarction. METHODS: Eighteen Chinese miniature pigs were modeled by the ligation of left anterior descending coronary artery. BrdU-labeled bone marrow mesenchymal stem cells were injected via the coronary artery at 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks after modeling, respectively. Then, the animals were killed at 3 days after cell transplantation to detect the home of bone marrow mesenchymal stem cells in the infarct zone. RESULTS AND CONCLUSION: BrdU-labeled positive cells with brown nuclei were visible at 1 day, 3 days, 1 week, 2 weeks, 3 weeks, and 4 weeks after myocardial infarction, especially at 1 week after myocardial infarction (P < 0.05). It indicates that the best homing window for bone marrow mesenchymal stem cells was at week after myocardial infarction, when the stem cell transplantation is given to be able to promote myocardial repair. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells regulate nuclear factor kappaB expression in alveolar macrophages of acute lung injury rats with sepsis Zhang Ji-feng, Zhang Zi-qi, Luo Xiao-tian, Hou Lin-yi, Jiang Qin, Lv Jie-ping, Zhang Wen-kai 2015, 19 (10):  1556-1561.  doi: 10.3969/j.issn.2095-4344.2015.10.015 Abstract ( 304 )   PDF (2313KB) ( 582 )   Save BACKGROUND: Bone marrow mesenchymal stem cells have a therapeutic effect on acute lung injury, but the mechanism is unclear. If the mechanism is understood, the majority of patients with acute lung injury can obtain a benefit.   OBJECTIVE: To explore the possible mechanism underlying bone marrow mesenchymal stem cells in the treatment of acute lung injury with sepsis in rats. METHODS: (1) Thirty-six adult Wistar rats were randomly divided into three groups, sham operation group (sham group), sepsis group and bone marrow mesenchymal stem cells group (cell treatment group). In the sepsis and cell treatment groups, animal models of sepsis with acute lung injury were established by cecal ligation and puncture, while in the sham group, the cecum was not ligated and punctured. Then, 1 mL normal saline was injected via the femoral vein in the sepsis and sham groups, and 1 mL bone marrow mesenchymal stem cell suspension (1×109/L) was injected into the cell treatment group. After 6 hours, interleukin 10 and macrophage inflammatory protein-2 levels in serum were measured in the three groups. Lung tissues were taken for pathological observation using hematoxylin-eosin staining. (2) Rat alveolar macrophages were obtained by bronchoalveolar lavage, seeded into 24-well culture plates, and divided into three groups: control group (group A), sepsis model group (group B) and intervention group of bone marrow mesenchymal stem cells (group C). Normal saline, septic plasma, and co-intervention of septic plasma and mesenchymal stem cells were used in the groups A, B, C, respectively. Then, cells in the three groups were cultured in a 5% CO2 incubator at 37 ℃ for 1 hour. After that, alveolar macrophages were taken to detect whether nuclear factor-κB (P65) protein entered into the nucleus using laser scanning confocal microscopy. RESULTS AND CONCLUSION: (1) The results of animal experiments showed that compared with the sham group, the macrophage inflammatory protein-2 levels in the sepsis group and cell treatment group were significantly increased (P < 0.05), but the macrophage inflammatory protein-2 level in the cell treatment group was significantly lower than that in the sepsis group (P < 0.05); there were no significant differences in serum interleukin 10 levels among the three groups (P > 0.05); inflammatory cell infiltration, interstitial pulmonary edema and pulmonary hemorrhage existed in the sepsis and cell treatment groups, but these symptoms were significantly reduced in the cell treatment group compared with the sepsis group. (2) Results from cell experiments showed that compared with the group A, in group B and group C, the number of nuclear factor-κB (P65) proteins into the nucleus was significantly higher (P < 0.05), but it was lower in the group C than the group B (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells in acute lung injury with sepsis can regulate nuclear factor-κB (P65) protein of alveolar macrophages into the nucleus, reduce expression of macrophage inflammatory protein-2, and thereby play a protective role in the lung via reducing neutrophil infiltration. Temporarily, this study cannot explain whether bone marrow mesenchymal stem cells have an effect on interleukin 10. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Quantitative comparison of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells   Wang Yan-lan, Wang Xiao-xiao, Chen Tian-yu, Yang Fan-qiao, Dai Xu-bin, Ou Qian-min, Dai Shuang-ye, Lin Xue-feng 2015, 19 (10):  1562-1569.  doi: 10.3969/j.issn.2095-4344.2015.10.016 Abstract ( 363 )   PDF (499KB) ( 575 )   Save BACKGROUND: Human periodontal ligament stem cells are a kind of mesenchymal stem cells that have self-renewal and multidifferentiation potential. Previous studies have showed that human periodontal ligament stem cells can differentiate into osteoblast-like cells or adipocyte-like cells under appropriate induction. Yet few  studies have focused on the expression level of parathyroid hormone 1 receptor which will affect the osteogenic potential of Human periodontal ligament stem cells.   OBJECTIVE: To examine the expression level of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells and to discuss the role of parathyroid hormone 1 receptor in osteogenic differentiation. METHODS: By using magnetic-bead cell sorting, we separated and identified the human periodontal ligament stem cells and human periodontal ligament cells. We examined and compared the mRNA expression level of parathyroid hormone 1 receptor in human periodontal ligament stem cells and human periodontal ligament cells by Real-Time PCR. Osteoblastic differentiation was examined through in vitro matrix mineralization by alizarin red staining and alkaline phosphatase assay. RESULTS AND CONCLUSION: Positive immunomagetic sorted cells were positive for STRO-1, CD146, Vimentin, indicating that they were periodontal ligament stem cells. Parathyroid hormone 1 receptor was expressed in human periodontal ligament stem cells and mainly located in cell membrane and cytoplasm which were similar to human periodontal ligament cells and MG63 cells. The expression of parathyroid hormone 1 receptor in human periodontal ligament stem cells was 3.7 times higher than that in human periodontal ligament cells, which was similar to that in MG63 cells. After osteogenic induction, human periodontal ligament stem cells showed a higher expression of parathyroid hormone 1 receptor and osteoblast-related genes as well as the activity of osteoblast alkaline phosphatase and mineralization compared to human periodontal ligament cells. Our data showed that parathyroid hormone 1 receptor was higher in human periodontal ligament stem cells than human periodontal ligament cells and the expression was related with osteogenic differentiation, suggesting that human periodontal ligament stem cells display a higher potency of osteogenic differentiation and act as seed cells with a vast application prospect in oral 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Tissue-engineered tubular grafts constructed with stem cells and bladder acellular matrix in long-segmental ureteral reconstruction   Liao Wen-biao, Yang Si-xing, Xiong Yun-he, Song Chao, Meng Ling-chao 2015, 19 (10):  1570-1574.  doi: 10.3969/j.issn.2095-4344.2015.10.017 Abstract ( 280 )   PDF (1596KB) ( 335 )   Save BACKGROUND: The main way for long-segmental ureteral reconstruction may cause a lot of traumas and complications. Therefore, to seek a new repair method is urgent. OBJECTIVE: To investigate the feasibility of a tissue-engineered tubular graft for ureteral reconstruction. METHODS: Bone marrow mesenchymal stem cells and smooth muscle cells of rabbits were seeded into the two surfaces of bladder acellular matrix and cultivated for 7 days. Then the graft was used to prepare a 4-cm long tissue-engineered tubular graft, which was regarded as experimental group. Smooth muscle cells seeded onto the bladder acellular matrix was used to construct the tissue-engineered tubular graft as control group. Twenty-five New Zealand rabbits were randomly divided into experimental group (n=20) and control group (n=5), and two kinds of tubular grafts covered with omentum were implanted into the two groups, respectively, for repair of ureteral defects. Hematoxylin-eosin staining and immunohistochemical detection were performed at 2, 4, 8 weeks after implantation.  RESULTS AND CONCLUSION: In the experimental group, hematoxylin-eosin staining showed epithelial coverage and muscle fibers on the lumen of tissue-engineered tubular grafts at 8 weeks after implantation; immunohistochemistry showed that anti-AE1/AE3 antibody and anti-uroplakin Ⅲa antibody were positive, confirming that there were mature epithelial cells on the lumen of tissue-engineered tubular grafts. In the control group, five rabbits were dead within 2 weeks after removal of ureteral scaffold, and autopsy showed scar formation inside the graft and severe hydronephrosis. These results demonstrate that it is feasible to construct the tissue-engineered tubular graft using bone marrow mesenchymal stem cells and smooth muscle cells into the bladder acellular matrix for ureteral reconstruction. Bone marrow mesenchymal stem cells can potentially promote urothelial regeneration. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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DCLK1+/Ki67- cell morphology and distribution in colorectal cancer   Wang Huan, Ma Fa-ku, Liu Bin, Shi Min, Xiao Wei-ling 2015, 19 (10):  1575-1579.  doi: 10.3969/j.issn.2095-4344.2015.10.018 Abstract ( 459 )   PDF (389KB) ( 691 )   Save BACKGROUND: DCLK1 is a transmembrane microtubule-associated kinase in neurons after mitotic division, which may be the intestinal cancer stem cell marker. OBJECTIVE:To observe the expression and pathological significance of DCLK1 and Ki67 in colorectal cancer. METHODS: Expression of Ki67 and DCLK1 in 150 cases of colorectal cancer tissues was detected by immunohistochemical method in contrast to normal colorectal mucosa, para-carcinoma tissue, and adenoma tissue. RESULTS AND CONCLUSION: The expression rates of DCLK1 and Ki67 were 36.7% and 34.7% in cancer tissues, respectively, both of which were significantly higher than those in normal colorectal mucosa and adenoma. The expression of DCLK1 was associated with the location, depth of invasion, lymph node metastasis (P < 0.05), while the expression of Ki67 was just associated with the depth of invasion (P < 0.05). There was a negative correlation between the expression of DCLK1 and Ki67 (r=-0.460, P=0.000). The count of DCLK1+/Ki67-cells was about 2.01% in colorectal cancer tissues, and these cells mainly distributed at the bottom of intestinal mucosa base and common duct wall. DCLK1+/Ki67- cells were oval, the nuclei were large and deep-stained with prominent nucleolus, and there was rare nuclear fission and less cytoplasm. From the aspects of cell number, location, and cell morphology, DCLK1+/Ki67- cells are in line with the characteristics of cancer stem cells; therefore, DCLK1+/Ki67- can be used as a cancer stem cell marker of colorectal cancer. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Co-culture of ginsenosides Rg1 and neural stem cells: promoting proliferation role and protective effect Sun Jian-zhong, Liu Xin-wei, Guan Hua-peng, Zhang Peng, Liu Qi, Yang Jun, Guo Qun-feng, Ni Bin 2015, 19 (10):  1580-1584.  doi: 10.3969/j.issn.2095-4344.2015.10.019 Abstract ( 284 )   PDF (342KB) ( 532 )   Save BACKGROUND: Chinese herb extracts can restore and protect the nervous system of rats through intervention of neural stem cells. OBJECTIVE: To explore the role of ginsenosides Rg1 in the proliferation and protection of neural stem cells. METHOD: Sprague-Dawley rats at pregnant 19 days were dissected to take out fetal rats, and then the hippocampal tissues from fetal rats were isolated to extract neural stem cells. Neural stem cells were co-cultured with DMEM/F12 medium containing 50 g/L ginsenosides Rg1 as intervention group, with DMEM/F12 medium as blank control group, and with DMEM/F12 containing 0.64% phenol as positive control group, respectively. MTT assay was used to detect the proliferation of neural stem cells in each group, and western blot method to detect the protein expression of brain-derived neurotrophic factor and transforming growth factor-β in neural stem cells. RESULTS AND CONCLUSION: Rat neural stem cells were round single cells with clear border at early period after isolation but at 2 days after inoculation, the cells were adherent and aggregated into small cell spheres. Compared with the blank control group, the proliferative rate of neural stem cells was significantly increased in the ginsenosides Rg1 group (P < 0.05), but decreased in the positive control group (P < 0.05). Compared with the blank control group, in the ginsenosides Rg1 group, the expression of brain-derived neurotrophic factor was elevated, and the expression of transforming growth factor-β was reduced, indicating ginsenosides Rg1 has a certain effect to promote the proliferation of neural stem cells as well as to protect the neural stem cells. 1库： 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Mesenchymal stem cells derived from adipose and nucleus pulposus tissue differentiate towards nucleus pulposus-like cells induced by transforming growth factor-beta 1 Xue Chen-hui, Ma Xun, Guan Xiao-ming, Zhang Hui, Zhang Li 2015, 19 (10):  1585-1891.  doi: 10.3969/j.issn.2095-4344.2015.10.020 Abstract ( 239 )   PDF (473KB) ( 451 )   Save BACKGROUND: A large number of studies have shown that adult stem cells derived from multiple tissues are available to differentiate towards nucleus pulposus-like cells in vitro. It is unclear whether mesenchymal stem cells derived from nucleus pulposus tissues have the ability to differentiate towards nucleus pulposus-like phenotypes induced by transforming growth factor-beta 1. Up to now, there are few reports on the difference between the differentiation ability of mesenchymal stem cells derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cells.   OBJECTIVE: To compare the ability of mesenchymal stem cells derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cells differentiating into nucleus pulposus-like cells under induction of transforming growth factor-beta 1. METHODS: The groin fat tissue and the coccygeal spine of rats were taken respectively to isolate and culture mesenchymal stem cells derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cells by mechanical enzyme digestion method. Flow cytometry was employed to detect the expression of CD105, CD90, CD29, CD45, CD44, CD34, and CD24 of both two kinds of stem cells. Mesenchymal stem cells derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cells were divided into complete induction group (complete induction medium with transforming growth factor-beta 1), incomplete induction group (complete induction medium without transforming growth factor-beta 1) and control group(DMEM/F12 containing 10% fetal bovine serum and 100 mg/L penicillin/streptomycin), respectively. After 14 days of culture, real-time PCR was used to detect the expression of collagen type II, Aggrecan and SOX-9 in each group. RESULTS AND CONCLUSION: CD105, CD90, CD29 expressed positively and CD45, CD44, CD34, CD24 negatively in both two kinds of stem cells. After 14 days of induced differentiation, the expressions of collagen type II, Aggrecan and SOX-9 in the two kinds of cells were significantly higher in the complete induction groups than in the control groups (P < 0.05). Under the induction of transforming growth factor-beta 1, the expression of collagen type II, Aggrecan and SOX-9 in mesenchymal stem cells derived from nucleus pulposus tissues was significantly higher than that in adipose-derived mesenchymal stem cells (P < 0.05). These findings suggest that both two kinds of mesenchymal stem cells have the ability to differentiate towards nucleus pulposus-like cells induced by transforming growth factor-beta, and mesenchymal stem cells derived from nucleus pulposus tissues may be more suitable as seed cells for nucleus pulposus tissue engineering research. 1库： 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Hypoxia-inducible factor-1alpha gene-modified umbilical cord blood mesenchymal stem cells transplantation in the treatment of spinal cord ischemia-reperfusion injury   Fan Zhi-gang, Shi Rui-hong 2015, 19 (10):  1592-1596.  doi: 10.3969/j.issn.2095-4344.2015.10.021 Abstract ( 187 )   PDF (830KB) ( 415 )   Save BACKGROUND: There are no available therapies for spinal cord ischemia-reperfusion injury, and stem cell transplantation is a focused topics. OBJECTIVE: To observe the therapeutic effect of hypoxia-inducible facotr-1α gene-modified umbilical cord blood mesenchymal stem cells (UC-MSCs) transplanted via the infrarenal abdominal aorta on spinal cord ischemia-reperfusion injury in rats. METHODS: Thirty adult female Sprague-Dawley rats were randomly divided into three groups, with 10 rats in each group. The infrarenal abdominal aorta of rats was occluded surgically for 1 hour, and then the spinal cord reperfusion was restored. At 2 hours after reperfusion, 1 mL of 10% PBS, UC-MSCs suspension and hypoxia-inducible factor-1α-modified UC-MSCs suspension was injected via the infrarenal abdominal aorta, respectively, in the three groups. At 1, 6, 12 days after injection, Basso-Beattie-Bresnahan scores were recorded and western blot assay was used to detect hypoxia-inducible factor-1α protein expression in the spinal cord. The motor-evoked potential was determined at 12 days after injection. RESULTS AND CONCLUSION: Compared with the control group, the Basso-Beattie-Bresnahan scores were significantly higher (P < 0.05), the expression of hypoxia-inducible factor-1α protein in the spinal cord tissue was significantly increased (P < 0.05), the motor-evoked potential latency was shortened (P < 0.05) and the amplitude was increased (P < 0 .05) in the untransfected group and transfection group. Compared with the untransfected group, the Basso-Beattie-Bresnahan scores were significantly higher (P < 0.05), the expression of hypoxia-inducible factor-1α protein in the spinal cord tissue was significantly increased (P < 0.05), the motor-evoked potential latency was shortened (P < 0.05) and the amplitude was increased (P < 0 .05) in the transfection group. Above all, umbilical cord blood mesenchymal stem cell transplantation modified by hypoxia-inducible factor 1α has better effects on spinal cord ischemia-reperfusion injury. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation for treatment of cis-platinum-induced ovarian damage Ye Xiao-feng, He Yuan-li, Fu Xia-fei, Wang Xue-feng 2015, 19 (10):  1597-1602.  doi: 10.3969/j.issn.2095-4344.2015.10.022 Abstract ( 384 )   PDF (844KB) ( 429 )   Save BACKGROUND: Chemotherapy drugs can damage the ovarian function in women of childbearing age, and even lead to premature ovarian failure. Therefore, to improve and restore the ovarian function in patients has become an important issue. OBJECTIVE: To explore the therapeutic effect and feasibility of bone marrow mesenchymal stem cell therapy against chemotherapy-induced ovarian damage． METHODS: Rat models of chemotherapy-induced premature ovarian failure were established, and injected with PKH26-labeled bone marrow mesenchymal stem cells. At 15, 30, 45, 60 days after cell transplantation, five rats were selected respectively to detect follicle-stimulating hormone and estradiol levels, and then, the rats were killed to take the right ovary for pathological examination. The number of ovarian follicles was detected under light microscope. At 30 days after cell transplantation, another two rats were selected to mate with male rats to observe the difference in the reproductive activity. RESULTS AND CONCLUSION: Four of 22 rats (18%) gradually recovered their estrous cycle after cell  transplantation, with the decreased follicle-stimulating hormone level and increased estradiol level. Moreover, the number of follicles was reduced. All of these indicated that the ability to have children in rats was not damaged. These experimental findings suggest that bone marrow mesenchymal stem cells can partially improve the ovarian function of rats under chemotherapy. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Autologous hematopoietic stem cell transplantation for peripheral T cell lymphoma Pan Yao-zhu, Bai Hai, Wang Cun-bang, Xi Rui, Zhang Qian, Wang Xiao-jing 2015, 19 (10):  1603-1607.  doi: 10.3969/j.issn.2095-4344.2015.10.023 Abstract ( 340 )   PDF (519KB) ( 563 )   Save BACKGROUND: The incidence rate of peripheral T cell lymphoma is high in Asia, and peripheral T cell lymphoma is aggressive with generally poor prognosis. However, there is no standard treatment strategy.   OBJECTIVE: To retrospectively analyze the therapeutic effect of autologous hematopoietic stem cell transplantation on peripheral T cell lymphoma as well as relevant toxic and side effects. METHODS: A retrospective review was conducted in 35 patients with peripheral T cell lymphoma who underwent autologous hematopoietic stem cell transplantation from March 2003 to April 2014, including 22 cases of extranodal NK/T-cell lymphoma (nasal type), 1 case of angioimmunoblastic T-cell lymphoma, 8 cases of peripheral T cell lymphoma (non-specific), 3 cases of ALK-positive anaplastic large cell lymphoma, and 1 case of ALK-negative anaplastic large cell lymphoma. All of 35 patients were classified pathologically according to WHO pathological type in 2001 and 2008, and received the high-dose chemotherapy with vincristine, cytarabine, etoposide, mitoxantrone, semustine, cyclophosphamide, and total body irradiation. RESULTS AND CONCLUSION: After a median follow-up of 54 (9-120) months, the probabilities of overall survival and disease-free survival after transplantation were 80% (n=28) and 71% (n=25), respectively. Eight cases (23%) relapsed after transplantation, seven of which died. It was safe with mild and moderate transplantation related side-effects on opportunistic infections, oral cavity mucosa and bladder responses and so on, and there were no severe, life-threatening late complications. Autologous hematopoietic stem cell transplantation may be an effective and safe treatment for peripheral T cell lymphoma, and there is a better benefit in peripheral T cell lymphoma patients with first complete remission． 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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A new method to isolate mesenchymal stem cells from human placenta Liu Yang, Li Yan-qi, Wang Hong-yi, Wu Xiao-bing, Jing Yong-guang, Xu Xiao, Yao Yao, Zhang Yu, Chu-Tse Wu, Jin Ji-de 2015, 19 (10):  1608-1612.  doi: 10.3969/j.issn.2095-4344.2015.10.024 Abstract ( 620 )   PDF (832KB) ( 1010 )   Save BACKGROUND: Placenta is a valuable source of mesenchymal stem cells for stem cell therapy and future application in the field of regenerative medicine. However, conventional methods cannot acquire a large amount of purified human placenta-derived mesenchymal stem cells. Here, we present a new method for isolating human placenta-derived mesenchymal stem cells suitable for banking strategies and for future clinical applications. OBJECTIVE: To analyze the biological characteristics of human placenta-derived mesenchymal stem cells cultured by tissue dissociating and collagenase digestion. METHODS: Human placenta-derived mesenchymal stem cells were obtained from human placenta by tissue dissociating and collagenase digestion method. Immunophenotype was analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated by in vitro adipogenic, osteogenic and chondrogenic induction as well. RESULTS AND CONCLUSION: Human placenta-derived mesenchymal stem cells could be passaged stably in vitro. Furthermore, the cells expressed CD73, CD90, CD105, but were negative for the markers of CD11b, CD19, CD34, CD45, and HLA-DR. Human placenta-derived mesenchymal stem cells proliferated actively and began to grow logarithmically at days 3-5 followed by a plateau period at day 6. In addition, the isolated cells could be induced into adipocytes, osteocytes, chondrocytes in vitro. In a word, the results of this study demonstrated that the tissue dissociating and collagenase digestion method is an efficient method for obtaining a large amount of human placenta-derived mesenchymal stem cells that can be stably cultured in vitro and have strong proliferative ability. 1库： 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell culture technology: innovation, breakthrough and high quality Tu Xue-song 2015, 19 (10):  1613-1618.  doi: 10.3969/j.issn.2095-4344.2015.10.025 Abstract ( 287 )   PDF (360KB) ( 420 )   Save BACKGROUND: With the development of fundamental research and clinical research on stem cells, the quantity demanded for stem cells is on the increase, but the relatively out-dated cultivating technology of stem cells restrict the development of the research on stem cells.   OBJECTIVE: To review the improvement and innovation of stem cell culture techniques both at home and abroad. METHDOS: In the title and abstract, “stem cells, induced pluripotent stem cells, cultivation, efficiency, reprogramming” in Chinese and English chosen as search terms were utilized to search for the papers related to stem cell culture techniques published from January 2005 to November 2014 in CNKI, VIP, Wanfang and PubMed databases, respectively. In the same field, articles published lately in the authoritative journals were preferred. Totally 61 articles were searched initially, and only 39 articles were included in result analysis. RESULTS AND CONCLUSION: The development and improvement of stem cell culture techniques effectively increase the culturing quantity of stem cells, improve the development of basic and clinical studies on stem cells. With the further innovation and breakthrough of stem cell culture techniques, it will be possible to efficiently culture high-quality stem cells, and the stem cell transplantation tests for human diseases will be promoted 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Induced pluripotent stem cell therapy: transplant rejection and safety Wu Peng-fei, Zhou Guang-ji 2015, 19 (10):  1619-1623.  doi: 10.3969/j.issn.2095-4344.2015.10.026 Abstract ( 257 )   PDF (273KB) ( 511 )   Save BACKGROUND: Induced pluripotent stem cells have been a hotspot in regenerative medicine research since it was discovered. The clinical application of induced pluripotent stem cells is excessively focused on, but the safety issue is almost ignored. OBJECTIVE: By summarizing the application of induced pluripotent stem cells in animal experiments to analyze the safety problems of induced pluripotent stem cells and their possible reasons in order to lay a foundation for further study and clinical application of induced pluripotent stem cells. METHODS: PubMed database was retrieved by the first author for articles related to the safety of induced pluripotent stem cells published from 2006 to 2014 using the keywords of “induced pluripotent stem cells, safety, immune, immunogenicity, tumorigenicity, cancer, epigenomic, transplantation, generation, reprogramming,genomic, mutation” in English. Related full texts were got from Cell Press and Nature Databases. Finally, 28 articles were chosen in result analysis. RESULTS AND CONCLUSION: Safety problems of induced pluripotent stem cells are attracting more and more attentions. Immunogenicity, potential tumorigenicity and epigenetic variation are major risks for the clinical applications of induced pluripotent stem cells. Safety issues of induced pluripotent stem cells mainly come from the reprogramming process. The “integrating genetic manipulation” may lead to a greater risk of tumorigenicity than non-integrating operations. Epigenetic variations emerge in the reprogramming, which are mostly relative to “epigenetic memory” of reprogrammed 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Poultry mesenchymal stem cells: exploration and prospects Li Dong-sheng, Wang Bing-yun,Chen Zhi-sheng, Chen Sheng-feng, Ji Hui-qin, Xian Qiong-zhen, Chen Jin-ding 2015, 19 (10):  1624-1629.  doi: 10.3969/j.issn.2095-4344.2015.10.027 Abstract ( 230 )   PDF (428KB) ( 378 )   Save BACKGROUND: Poultry mesenchymal stem cells are a particular subset of pluripotent adult stem cells derived from the mesoderm, which have great application prospects because of their strong proliferation and multi-directional differentiation potential. OBJECTIVE: To review the source, separation, purification, culture and differentiation of poultry mesenchymal stem cells, and to provide the theoretical foundation and experimental basis for the further research and application of poultry mesenchymal stem cells. METHODS: PubMed and CNKI databases were searched by the first author using key words of “mesenchymal stem cells, poultry, chicken, isolation, culture, differentiation” in English and in Chinese, respectively, to retrieve relevant articles published from 1990 to 2014. Literatures addressing induced poultry mesenchymal stem cells were included, and 42 articles were chosen for further analysis eventually. RESULTS AND CONCLUSION: Poultry mesenchymal stem cells have great application prospects in the aspects of establishing in vitro model of poultry cells, studying poultry disease pathogenesis, animal nutrition and meat quality control. Its origin source is wide and easy to obtain. Isolation, purification, culture and biological characteristics of mesenchymal stem cells from different tissues are different. But, the study on poultry mesenchymal stem cells is still in the exploration process, and there are many technical problems to be solved. 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Noninfectious pulmonary complications following hematopoietic stem cell transplantation Luo Hong, Zhang Hong-bin 2015, 19 (10):  1630-1634.  doi: 10.3969/j.issn.2095-4344.2015.10.028 Abstract ( 453 )   PDF (366KB) ( 487 )   Save BACKGROUND: Hematopoietic stem cell transplantation is a promising treatment for a variety of malignant and nonmalignant disorders. But noninfectious pulmonary complications following hematopoietic stem cell transplantation are a major cause of morbidity and mortality in these patients. And the diagnosis and treatment are difficult. OBJECTIVE: To review the noninfectious pulmonary complications after hematopoietic stem cell transplantation in terms of onset time, risk factors, clinical manifestations, characteristics of the high-resolution CT, histopathological measurement, related genes and treatment options. METHODS: A computer-based search of VIP, PubMed and Sciencedirect databases was performed for articles related to noninfectious pulmonary compli cations after hematopoietic stem cell transplantation published from January 2005 to October 2014. The key words were “HSCT, pulmonary complications, non-infectious, pirfenidone” in Chinese and English in the title and abstract. Finally, 31 articles were included in result analysis. RESULTS AND CONCLUSION: The incidence of noninfectious pulmonary complications after hematopoietic stem cell transplantation has become more and more, with atypical clinical manifestations and limited diagnosis and treatments. According to the different clinical onset time, diagnostic criteria, clinical manifestations and the appropriate laboratory tests, clinicians can make early diagnosis and early intervention. Especially, the usage of high-resolution chest CT and bronchofiberscope for bronchoalveolar lavage as well as the timely drug administration can improve the survival rate of patients with noninfectious pulmonary complications after hematopoietic 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 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell therapy for degenerative disc diseases: present status and prospects Zhou Yi-chi, Li Jing-feng, Dong Shi-shi, Jin Wei 2015, 19 (10):  1635-1639.  doi: 10.3969/j.issn.2095-4344.2015.10.029 Abstract ( 278 )   PDF (415KB) ( 502 )   Save BACKGROUND: None of the current treatment strategies has been focused on relieving or reversing the disk degeneration process after degenerative disc diseases. In recent years, more and more scientists try to treat degenerative disc diseases using stem cell therapy.   OBJECTIVE: To explore the research status and prospects of stem cell therapy for degenerative disc diseases. METHODS: A computer-based online search of PubMed database between January 2004 and December 2014was performed to search related articles with the key words of “stem cell, intervertebral disk” in English. Literatures related to stem cell therapy for degenerative disc diseases were selected; in the same field, the articles published lately in authoritative journals were preferred. RESULTS AND CONCLUSION: A total of 342 articles were primarily selected, and 43 articles were involved in result analysis according to inclusion criteria. Stem cell therapy is a newly treatment for degenerative disc diseases. Cells appropriate for stem cell therapy include embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, human umbilical cord mesenchymal stem cells and chondrocytes or nucleus pulposus cells. Although cell leakage, intervertebral disc infection and tumorigenesis are the main challenges, stem cell therapy for degenerative disc diseases is promising in the future. 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 全文链接： Figures and Tables | References | Related Articles | Metrics