乳鼠心肌膜微血管内皮细胞的体外培养

THE CULTURE OF MYOCARDIAL MICROVASCULAR ENDOTHELIAL CELLS OF SUCKLING RATS IN VITRO

目的改进心肌膜微血管内皮细胞的体外培养方法,为进一步研究其结构和功能提供实验数据。方法选取10-15d龄的SD大鼠,采用植块法培养原代心肌膜微血管内皮细胞,在继代培养中通过差速消化和差速贴壁方法、结合倒置显微镜下形态学的观察进行继代培养和纯化,利用免疫细胞化学方法检测第Ⅷ因子相关抗原对培养细胞进行了鉴定。结果成功培养了大鼠心肌膜微血管内皮细胞,细胞呈多边形或鹅卵石状;免疫细胞化学染色第Ⅷ因子相关抗原显阳性。结论改良了心肌膜微血管内皮细胞的体外培养方法,获得了较高纯度的心肌膜微血管内皮细胞,可用于进一步的科学研究。

Objective To improve methods of culturing rat myocardial microvascular endothelial cells （RMMVECs） in vitro and provide experimental data for studying the structures and functions of RMMVECs. Methods SD rats of 10-15 days old were selested. Explants cultivation of cardiac tissues was used to obtain the primary cells. In the subculturing, the cell cultures were purified by different speeds of attachment and digestion from contaminant cells combined with morphological observation. The purified cells were identified as endothelial cells by morphology and by positive immunocytochemistry for factor Ⅷ-related antigen, a marker for endothelial cells. Results The RMMVECs were cultured and purified successfully which grew in a polygonal or cobblestone pattern and were positive for the immunocytochemical staining of factor Ⅷ-related antigen. Conclusion The methods of culturing RMMVECs were improved, RMMVECs with high purity were obtained and available data were provided for the further studies.