目的探讨糖尿病大鼠冠状动脉平滑肌细胞体外培养方法,建立糖尿病大鼠冠状动脉平滑肌细胞模型,为糖尿病性冠心病的研究奠定基础。方法建立糖尿病大鼠模型,用酶消化法体外培养冠状动脉血管平滑肌细胞。结果糖尿病大鼠造模成功后分离冠状动脉,用酶消化法培养糖尿病大鼠血管平滑肌细胞,24h更换培养液液,培养7~10d细胞重叠生长达多层,高低起伏呈“峰-谷”状。细胞α-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。
Aim To investigate the method of culturing vascular smooth muscle cells (VSMC) of coronary arteries derived from experimental diabetic rats and to establish the model of VSMC for the study of chronic cardiovascular diseases of diabetes. Method After the establishment of the model of diabetic rats, the coronary arteries of rats were 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 isolated by different kinds of digestive enzyme in three steps. 7 - 10 days afterward, the cells overlapped in layers and displayed the typical ‘ hill and valley' pattern and positive immunochemical 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.