糖尿病大鼠血管平滑肌细胞体外培养方法的探讨

Study of the culture of vascular smooth muscle cells in vitro derived from experimental diabetic rats

目的:对糖尿病大鼠血管平滑肌细胞体外培养方法进行探讨,建立糖尿病大鼠血管平滑肌细胞模型,为糖尿病慢性血管并发症研究奠定基础。方法:建立糖尿病大鼠模型,用组织贴壁法体外培养胸主动脉血管平滑肌细胞。结果:糖尿病大鼠造模成功。用贴壁法培养糖尿病大鼠血管平滑肌细胞,3d后相差显微镜下可见细胞游出,培养7～10d左右细胞重叠生长达多层,高低起伏呈“峰—谷”状。平滑肌细胞actin免疫组织化学染色鉴定为平滑肌细胞。结论:糖尿病大鼠血管平滑肌细胞增殖速度快,培养条件要求严格,在形态学上与正常大鼠平滑肌细胞相同。

Objective To investigate the method of culturing vascular smooth muscle cells (VSMC) in vitro derived from experimental diabetic rats and to establish the model of VSMC for the study of chronic vascular diseases of diabetes. Method After the establishment of the model of diabetic rats, the aortic of rats was separated carefully to culture VSMC using the method of explanting culture. Results The primary culture of VSMC was performed successfully by the method mentioned above. The cells could be seen from the small tissue pieces 3 days later. 7-10 days afterward, the cells overlapped in layers and displayed the typical ‘hill and valley’ pattern and possitive immunocyto-chemical staining for smooth muscle actin. Conclusion VSMC from experimental diabetes proliferates faster than normal VSMC and its cultivating condition is strictly limited. The morphology of diabetic VSMC is as same as that of normal rats.