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大鼠脑基底动脉平滑肌细胞培养及功能鉴定
  • 期刊名称:丘钦英,周家国,刘玉洁,关永源 大鼠脑基底动脉平滑肌细胞培养及功能鉴定 中国药理学通报
  • 时间:0
  • 分类:R-332[医药卫生] R322.121[医药卫生—人体解剖和组织胚胎学;医药卫生—基础医学]
  • 作者机构:[1]中山大学中山医学院药理学教研室,广东广州510080
  • 相关基金:国家自然科学基金资助项目(No30472021,30500616);广东省自然科学基金资助课题(No05300822);中山大学中山医学院青年教师科研启动基金.
  • 相关项目:他汀类药物防治高血压脑血管重构及其与氯通道的关系
中文摘要:

目的探讨大鼠脑基底动脉平滑肌细胞的培养方法,了解生长特性。方法用组织贴块法培养大鼠脑基底动脉平滑肌,用免疫细胞化学法鉴定细胞,用RF-5000荧光分光光度计观察细胞内Ca2+浓度的动态变化来检测其活性。结果培养5d后可见组织块周围有平滑肌细胞长出,2wk后可融合传代,经平滑肌特异性肌动蛋白α-actin鉴定,确定为平滑肌细胞。钙离子通道激动剂可诱导细胞Fura-2荧光比值(F340/F380)上升,细胞活性良好。结论组织贴块法是大鼠脑基底动脉平滑肌细胞培养简单、高效、经济的实验方法,为脑血管疾病发病机制的研究提供了理想的细胞模型。

英文摘要:

Aim To investigate the method of cell culture for smooth muscle cells from rat cerebral basilar artery and understand cells growth and biological characteristics.Methods The explant attached method was applied for cell culture of rat basilar artery smooth muscle cells(BASMCs).The cultured BASMCs were identified by immunocytochemical staining.The activities of cells were indicated by the dynamic changes of intracellular calcium concentration observed by RF-5 000 fluorospectro-photometer.Results BASMCs grew out of tissue blocks by 5 days,reached confluency could be subcultured after 2 weeks.Cultured cells were identified by intensely positive immunocytochemical staining to smooth muscle actin-specific.Introduction of calcium channel agonists induced significant increase in Fura-2 fluorescence ratio(F340/F380)and cells were in good condiction.Conclusion Explant attached method is simple,efficient and economic.It provides an ideal cell model for the study of pathogenesis of the cerebral vascular diseases.

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