文献类型：期刊文献

中文题名：Wistar大鼠BMSCs的分离与生物学特性分析

英文题名：Isolation and Biological Characteristics of Bone Marrow Mesenchymal Stem Cells Derived from Wistar Rats

作者：薛娜[1];伍志伟[1,2];刘丹[1];刘永琦[1,2];苏韫[1];颜春鲁[1]

第一作者：薛娜

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃省高校重大疾病分子医学与中医药防治省级重点实验室,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2017

卷号：39

期号：3

起止页码：324

中文期刊名：中国细胞生物学学报

外文期刊名：Chinese Journal of Cell Biology

收录：CSTPCD;;CSCD:【CSCD2017_2018】；

基金：甘肃省自然科学基金(批准号:1212RJ2A081);甘肃省省属普通本科高校基本科研业务费专项(批准号:2305136301)资助的课题~~

语种：中文

中文关键词：骨髓间充质干细胞;Wistar大鼠;成骨分化;成脂分化

外文关键词：bone mesenchymal stem cells;Wistar rat;adipogenic differentiation;osteogenic differentiation

摘要：该文建立了一种简便、高效的Wistar大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分离方法,为研究其生物学特性奠定了基础。采用全骨髓贴壁培养与密度梯度离心结合筛选法无菌分离Wistar大鼠BMSCs,常规传代和成骨、成脂诱导后,经显微观察、细胞计数、免疫印迹、q PCR和流式细胞术检测,分析细胞形态、增殖与分化特性。结果表明,分离细胞传至3代后细胞呈漩涡状生长、形态为长梭形,细胞生长曲线呈"S"形,表面分子CD29、CD34、CD44、CD45和CD90的阳性率分别为98.21%、0.78%、91.62%、0.93%和99.27%。成脂和成骨诱导后,过氧化物酶体增殖物激活受体-γ2(PPAR-γ2)、脂蛋白脂肪酶(LPL)、骨髓基质细胞核心结合因子α1(Runx2)和骨形态发生蛋白-2(BMP-2)表达水平显著升高,细胞内分别出现了大量的红色脂滴和钙化结节。该研究运用全骨髓贴壁培养与密度梯度离心结合筛选法成功分离了BMSCs,分离细胞具有向成骨和成脂细胞分化的潜能与骨髓间充质干细胞的生物学特性。
This work was aim to establish a simple, efficient and optimum isolation and culture method of bone marrow mesenchymal stem cells（BMSCs） derived from Wistar rat. This method would lay the foundation for the study of its biological characteristics. In this study, BMSCs-derived from Wistar rats were isolated by the method-combined of whole bone marrow adherent culture and density gradient centrifugation under the sterile environment. The cells which were cultured in routine conditions and the cells which were induced respectively by osteogenic fluid and adipogenic fluid were observed by inverted phase contrast microscope, and the characteristics of cell proliferation and differentiation were analysed by counting cell, Western blot, Real-time PCR and flow cytometry.The results showed that the isolated cells presented typical uniform swirling growth state and long spindle-shapedafter the third generation,the growth curve was for＂S＂shape,the positive rate of cell surface antigen markers CD29,CD34,CD44,CD45 and CD90 were 98.21%,0.78%,91.62%,0.93%and 99.27%,respectively.After adipogenic and osteogenic induction,protein levels of PPAR-γ2,LPL,Runx2 and BMP-2 were significantly increased,and large amounts of lipid droplets and calcium nodulesappeared in the cellsof different induction by the Oil Red O and the Alizarin Red staining,respectively.BMSCs-derived from Wistar rat were successfully isolated and culutured by the method of whole bone marrow adherent culture and density gradient centrifugation.The cells-isolated have the potential abilities of osteogenic and adipogenic differentiation and the biological properties of BMSCs.