| Rspo1基因转染间充质干细胞株的构建及生物学活性鉴定 |
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| 引用本文： | 陆婷,葛彦,邓云,陈伟,宋莉,曹莎莎,居颂文,居颂光. Rspo1基因转染间充质干细胞株的构建及生物学活性鉴定[J]. 中国血液流变学杂志, 2013, 0(4): 592-595,622 |
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| 作者姓名： | 陆婷 葛彦 邓云 陈伟 宋莉 曹莎莎 居颂文 居颂光 |
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| 作者单位： | [1]苏州大学医学部基础与生物科学学院免疫学系,江苏苏州215123 [2]江苏省干细胞研究重点实验室,江苏苏州215007 [3]中国人民解放军第100医院妇产科,江苏苏州215007 [4]苏州大学附属第二医院普外科,江苏苏州215004 [5]南京医科大学附属苏州医院消化疾病与营养研究中心,江苏苏州215008 |
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| 基金项目： | 国家自然科学基金青年科学基金资助项目（81000912,81373149,31200661）;江苏省自然科学基金资助项目（BK2012621）;“青蓝工程”资助项目（2010） |
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| 摘 要： | 目的：构建稳定表达Rspo1的基因转染间充质干细胞株。方法将人Rspo1全长cDNA重组入逆转录病毒载体pEGZ-Term，通过与辅助病毒载体共转染293T细胞，包装为具有感染力的完整重组病毒载体，收集培养上清感染间充质干细胞C3H10 T1/2细胞株，筛选获得G418抗性的基因转染细胞，通过RT-PCR与流式细胞术检测感染后C3H10 T1/2细胞Rspo1分子的表达，并采用细胞计数法观察其上清对SW480结肠癌细胞增殖能力的影响。结果成功构建pEGZ-Term/Rspo1逆转录病毒表达载体，获得了稳定表达人Rspo1的基因转染细胞株C3H10/Rspo1，该细胞株上清能促进SW480结肠癌细胞增殖。结论建立了稳定表达Rspo1的基因转染间充质干细胞株，为进一步利用Rspo1和间充质干细胞靶向作用于肠道干细胞救治肠上皮损伤的研究奠定了基础。
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| 关 键 词： | Rspo1 间充质干细胞 基因转染细胞 |
Construction of Rspo1 Gene Transducted Mesenchymal Stem Cell Line and the Identiifcation of Biological Activity |
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| LU Ting,GE Yan,DENG Yun,CHEN Wei,SONG Li,CAO Sha-sha,JU Song-wen,JU Song-guang. Construction of Rspo1 Gene Transducted Mesenchymal Stem Cell Line and the Identiifcation of Biological Activity[J]. Chinese Journal of Hemorheology, 2013, 0(4): 592-595,622 |
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| Authors: | LU Ting GE Yan DENG Yun CHEN Wei SONG Li CAO Sha-sha JU Song-wen JU Song-guang |
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| Affiliation: | 1.Dept of Immunology, School of Biology and Basic Medical Sciences, Medical College, Soochow University, Suzhou, Jiangsu, 215123, China; 2.Key Lab for Stem Cell Research of Jiangsu Province, Suzhou, Jiangsu, 215007, China; 3.Dept of Obs/Gyn, 100th Hospital of PLA, Suzhou, Jiangsu, 215007, China; 4.Dept of General Surgery, the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, 215004, China; 5.Digestive Diseases and Nutrition Research Center, AffiliatedSuzhou Hospital of Nanjing Medical University, Suzhou, Jiangsu, 215008, China) |
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| Abstract: | Objective To construct a gene transfected mesenchymal stem cell line that stably expressed Rspo1. Methods The full length human Rspo1 cDNA was subcloned into retroviral expressing vector pEGZ-Term. The recombinant plasmid together with its helper virus vector was cotransfected into the package cell 293T. The C3H10 T1/2 cells were infected with the supernatant of the transfected 293T cells, and then were selected with G418. The G418 resistant cells were harvested for screening their C3H10/Rspo1 expression by RT-PCR and flow cytometry. The biological effect of supernatant was analyzed by cell counting. Results The pEGZ-Term/Rspo1 retrovirus expressing vector was constructed successfully and a cell line, named C3H10/Rspo1, which sta-bly expressed Rspo1 was obtained and its supernatant could promote colon cancer cell line SW480 proliferation. Conclusion In our study, a stable gene transfected mesenchymal stem cell line that expressed Rspo1 was estab-lished. It provides a valuable tool to explore the novel strategy to rescues the intestinal epithelium damage. |
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| Keywords: | Rspo1 mesenchymal stem cell gene transfected cell line |
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