小鼠平滑肌祖细胞培养及其体内外迁移功能的研究  

In vitro culture and migration assay of mouse smooth muscle progenitor cells

作　　者：王晴[1] 李和权[1] 周建英[1] 

机构地区：[1]浙江大学医学院附属第一医院呼吸内科,浙江杭州310003

出　　处：《中国药理学通报》2016年第7期915-920,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助课题(No 81100020;81170038)

摘　　要：目的建立一种有效的小鼠平滑肌祖细胞的培养方法,并对其迁移功能进行研究。方法使用C57BL/6小鼠制备密质骨来源间充干细胞,PDGF-BB诱导其分化并采用形态学观察、免疫细胞化学染色及流式分析的方法进行鉴定。使用Transwell小室及流式分析方法检测其迁移能力。结果 PDGF-BB诱导7 d后,镜下可见细胞呈长梭型,免疫细胞化学染色显示开始表达α-SMA。诱导21 d后,流式分析证实70%以上的细胞表达CD34+/α-SMA+双阳性,58.5%细胞开始表达SM-MHC。迁移实验表明,培养得到的平滑肌祖细胞在体内外均具有良好的迁移能力。结论通过使用密质骨来源间充干细胞制备平滑肌祖细胞具有操作简便、获取率高及培养周期短等优点,培养得到的平滑肌祖细胞在体内外均具有迁移能力,适于进行后续功能及机制研究。Aim To establish a reliable method for the culture of mouse smooth muscle progenitor cells and in-vestigate their migration ability .Methods Mesenchy-mal stem cells from compact bones were obtained from C57 BL/6 mice and stimulated with PDGF-BB to in-duce these cells to differentiate into smooth muscle pro-genitor cells. Morphological analysis , immunocyto-chemical and flow cytometric analysis were used to i-dentify the cell type and the migration ability was in-vestigated by the transwell system and flow cytometry . Result After PDGF-BB stimulation for 7 days, the cells showed spindle shape and started to express α-SMA as demonstrated by immunocytochemistry .After＆nbsp;21 days induction , Flow cytometric analysis revealed that over 70%of the cells expressed both CD 34 andα-SMA and 58.5%of the cells expressed SM-MHC.Mi-gration assay showed that the smooth muscle progenitor cells from culture could migrate in vivo and in vitro. Conclusions The culture of smooth muscle progenitor cells from compact bone-derived mesenchymal stem cells is easily operated with high yield rate and shorten culture period . Obtained smooth muscle progenitor cells from culture could migrate in vivo and in vitro, which is suitable for the mechanism studies .

关 键 词：平滑肌祖细胞 间充干细胞 培养 迁移 平滑肌肌动蛋白 PDGF-BB 

分 类 号：R-332[医药卫生] R322.74