[人脐动脉血管平滑肌细胞体外成骨细胞分化和钙化]。

Yong Ping Guo, Ming Shu Sun, Jia Qi Qian, Zhao Hui Ni
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引用次数: 0

摘要

目的:研究人脐动脉血管平滑肌细胞能否在体外自发向成骨细胞分化。用MTT法获得了人脐动脉血管平滑肌细胞的生长曲线。形态学观察了VSMCs自发形成多细胞结节的过程。采用Hoechst 33258法和TUNEL法分别检测肿瘤中VSMCs的凋亡情况。免疫组化法检测碱性磷酸酶在结节组织中的表达。并分别用透射电镜和茜素红S对其钙化进行了研究。光镜下发现,传代7天后,脐动脉平滑肌细胞汇合,呈典型的“山谷”状。随着培养时间的延长,细胞在“山”区聚集形成结节。培养4-5周后,这些结节形成并自发钙化。碱性磷酸酶的表达与VSMCs的凋亡同时发生。实验结果表明,人脐动脉血管平滑肌细胞与主动脉血管平滑肌细胞一样,可以在体外自发地向成骨细胞分化,而细胞凋亡可能参与了这一过程。
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[Vascular smooth muscle cells from human umbilical artery undergo osteoblast differentiation and calcification in vitro].

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.

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