大鼠胸主动脉平滑肌细胞的培养与鉴定

目的探讨平滑肌细胞培养方法,了解生长特性。方法用组织贴块法培养大鼠胸主动脉平滑肌细胞,用相差显微镜观察其生长情况,用免疫荧光染色分析其抗原的表达。结果培养3日可见组织块周边有细胞长出,2周达亚融合,传代后细胞“谷峰”生长明显。SM-α-肌动蛋白和Calponin抗体免疫荧光染色呈阳性。结论组织贴块法是简单、经济、高效的平滑肌细胞培养方法,为心血管疾病发病机制的研究提供了理想的细胞模型。

Culture and identification of rat thoracic aorta smooth muscle cells

Objectives To establish a culture approach of smooth muscle cells and analyze cell growth and biological characteristics. Methods The thoracic aorta of rat was separated carefully and cut into small tissue pieces. These explants were seeded onto culture flasks and cultured in complete medium consisting of Dulbecco modified Eagle medium containing selected lots of 20% fetal calf serum and antibiotics at 37 in a humidified atmosphere with 5% carbon dioxide. Cells growth was observed under light microscope. Immunofluorescent stain was performed to analyze the expression of smooth muscle cell specific antigen. Results A few of cells grew out of tissue-block about 3 days late and approached confluence approximately 2 weeks. A typical " hill-valley" growth pattern was displayed after cells were passaged. Immunofluorescent stain revealed that 98% of the cultured cells was positive for SMa-actin and calponin. Conclusions The method of attachment-block culture of smooth muscle cells is simple and convenient. It provides a good cell model for the study of cardiovascular diseases.