体外培养的人脐动脉平滑肌细胞可自发成骨细胞分化并钙化  

VASCULAR SMOOTH MUSCLE CELLS FROM HUMAN UMBILICAL ARTERY UNDERGO OSTEOBLAST DIFFERENTIATION AND CALCIFICATION IN VITRO

作　　者：郭永平[1] 孙明姝[1] 钱家麒[1] 倪兆慧[1] 

机构地区：[1]上海交通大学医学院仁济医院肾脏科,上海200001

出　　处：《分子细胞生物学报》2008年第2期87-95,共9页Journal of Molecular Cell Biology

基　　金：国家自然科学基金(30370653);上海市教育委员会科研项目(04BC35)资助项目~~

摘　　要：研究脐动脉来源的平滑肌细胞在体外培养时是否会自发成骨细胞分化,以深入了解该细胞的生物学特点。采用MTT法研究了脐动脉平滑肌细胞的生长曲线;用形态学方法观察了脐动脉平滑肌细胞自发细胞结节的形成过程;用Hoechst 33258染色法及TUNEL染色法研究了细胞结节中的凋亡现象;采用免疫组化染色法研究了细胞结节中碱性磷酸酶的表达。采用茜素红S染色及透射电镜研究了脐动脉平滑肌细胞的自发钙化。研究发现,体外培养的脐动脉平滑肌细胞传代后约7天细胞即可汇合,汇合后的细胞表现为典型的"峰、谷"样生长状态,随着培养时间的延长,在"峰"形生长区域,细胞聚集生长,自发形成细胞结节,经4-5周培养,细胞结节不断增大并钙化,免疫组化发现结节中有大量碱性磷酸酶表达。同时,在结节形成过程中伴随有大量平滑肌细胞凋亡。我们的研究表明,体外培养的脐动脉来源的平滑肌细胞同主动脉来源的平滑肌细胞一样可以自发成骨细胞分化,而平滑肌细胞凋亡可能参与了该过程。To research if the vascular smooth muscle cells （VSMCs） from human umbilical artery undergo osteoblast differentiation spontaneously in vitro. The growth curve of vascular smooth muscle cells from human umbilical artery was obtained by MTT method. The course of multicell nodule formation spontaneously by VSMCs was observed morphologically. The apoptosis of VSMCs in the nodules was detected by Hoechst 33258 and TUNEL methods respectively. The expression of alkaline phosphotase in the nodules was detected by immunohistochemical method. And the calcification was studied with transmission electron microscope and by alizarin red S respectively. We found that the umbilical artery smooth muscle cells confluenced after 7 days of passage and exhibited typical ＂hill and valley＂ pattern under light microscope. The cells grew into aggregation and formed nodules at the ＂hill＂ region with culture-time prolongation. After 4-5 weeks culture, these nodules built up and calcified spontaneously. We also found alkaline phosphotase expression and apoptosis of VSMCs in these nodules at the same time. We conclude that the vascular smooth muscle cells from human umbilical artery just like from aortic artery can undergo osteoblast differentiation spontaneously in vitro, and apoptosis participate this procedure probably.

关 键 词：血管平滑肌细胞 成骨细胞分化 血管钙化 细胞培养 脐动脉 

分 类 号：Q813[生物学—生物工程]