心肌微血管内皮细胞培养及生物学特性的初步观察

目的 :建立一种新的心肌微血管内皮细胞培养方法 ,并初步观察其生物学特性。方法 :采用组织块干涸翻转法培养心肌微血管内皮细胞 ,Ⅷ因子相关抗原免疫组织化学鉴定 ,细胞计数法测定其生长曲线 ,3 H TdR掺入法及放射免疫法观察牵张刺激对其增殖能力和内皮素释放的影响。结果 :Ⅷ因子相关抗原鉴定表明心肌组织块培养的微血管内皮细胞纯度达 96 % ,其生长特点是潜伏期较心肌成纤维细胞长 ,达 72h ,在不加内皮细胞生长支持物时 ,传代 3～ 5次后 ,其生长速度逐渐降低。牵张刺激可显著诱导其增殖和释放内皮素。结论 :心肌微血管内皮细胞组织块培养法简便易行 ;其生长潜伏期较长 ,长期培养时需要内皮细胞生长因子 ;机械刺激可促进其生长和内分泌活化。

A Method for Cardiac Microvascular Endothelial Cell Culture and Preliminary Study of Its Biological Characters

Objective: To establish a new method for cardiac microvascular endothelial cell culture and preliminarily observe its biological character. Method: Ventricle of rats were cut into fine pieces and cultured by method of adhesion to petri plates, the cardiac microvascular endothelial cells were determined by Ⅷ factor-related antigen immunohistochemistry. Growth curve was measured by cell count, proliferation potential and endothelin release stimulated by stretch were estimated by 3H-TdR incorporation and radioimmunoassay. Results: Cells cultured were typical form of endothelial cells and the cell purity was 96% tested by immunohistochemistry of Ⅷ factor-related antigen. Itscharacterisation of growth was that incubation period was 72 hours and longer than that of cultured cardiac fibroblasts, and its growth velocity is gradually decreased after 3 to 5 passages when without endothelial cell supportant. The 3H-TdR incorporation and endothelin release were elevated significantly stimulated by stretch. Conclusion: The method was simple and convenient, the incubation period of cultured cardiac microvascular endothelial cells was longer, and its proliferation and endocrine were activated by stretch.