CAJ | 학술논문

目的探讨肺微血管内皮细胞(PMVECs)的分离培养方法。方法应用复合酶消化法获取Sprague Dawley大鼠的PMVECs，经传代培养纯化，并通过观察光镜下细胞的形态学特征、免疫细胞化学染色检测细胞内Ⅷ因子抗原的存在、透射电镜观察细胞超微结构等进行综合鉴定。结果体外培养的原代PMVECs在光镜下呈典型的铺路石样排列，细胞生长状态良好，抗Ⅷ因子相关抗原染色阳性。透射电镜可见Weibel-Palade小体，上述特征证实所获得细胞为PMVECs。结论复合酶消化法获取PMVECs的成功率和纯度高，细胞活力好，数量大，同时保有了PMVECs的结构和功能，可用于进一步的实验研究。
목적탐토폐미혈관내피세포(PMVECs)적분리배양방법。방법응용복합매소화법획취Sprague Dawley대서적PMVECs，경전대배양순화，병통과관찰광경하세포적형태학특정、면역세포화학염색검측세포내Ⅷ인자항원적존재、투사전경관찰세포초미결구등진행종합감정。결과체외배양적원대PMVECs재광경하정전형적포로석양배렬，세포생장상태량호，항Ⅷ인자상관항원염색양성。투사전경가견Weibel-Palade소체，상술특정증실소획득세포위PMVECs。결론 복합매소화법획취PMVECs적성공솔화순도고，세포활력호，수량대，동시보유료PMVECs적결구화공능，가용우진일보적실험연구。
Objective To investigate the method for isolation and culture of pulmonary microvascular endothelial cells (PMVECs). Methods Primary PMVECs were obtained by complex phosphoesterasum digesting from isolated lung tissues of SD rats. PMVECs were purified by serial subcultivation and identified by morphological characteristics by light microscope, the expression of Ⅷ factor (vWF) detected by immunocytochemical staining, and ultrastructure characteristics under transmission electronic microscopy. Results The primiary PMVECs presented a typical cobblestone morphology and in good condition. vWF was expressed in cytoplasm of the purified cells. Weibel-Palade bodies were shown by transmission electronic microscopy. These features confirmed the cells obtained were PMVECs. Conclusions Methods for isolation and culture of PMVECs by using complex phosphoesterasum digesting in our research can acquire high quantity and viable cells,which maintain the structure and function of PMVECs and can be used in subsequent research.