摘要
目的探索和建立一种高效的人脐动脉内皮细胞分离和培养方法。方法用PBS灌注清洗脐动脉后,以0.1 g/L1型胶原酶消化脐动脉内膜,收集、离心消化液,重悬细胞在特定培养基中培养。观察其形态特点,同时用CD31免疫荧光染色和内皮细胞管状结构形成实验对所得细胞进行鉴定。结果倒置相差显微镜下观察所获得的细胞为单层生长,呈现铺路石样形态,并且大量表达内皮细胞特异性膜蛋白CD31,同时能够形成明显的管状结构。结论本研究建立了操作简单、快速、高效分离人脐动脉内皮细胞的方法,所分离得到的内皮细胞纯度高、成活率高。
Objective To explore and establish an efficient system for isolation and culture of human umbilical arterial endothelial cells （HUAECs）. Methods The human umbilical arteries were perfused and washed by PBS, and then digested by 0.1g/L type 1 collagenase. Digested cell supernatants were collected, centrifuged, and the re-suspended cells were cultured in a specific medium. The identification of endothelial cells was carried out by observing the cellular morphology, immunostaining the endothelial surface marker CD31 under fluorescent microscopy and examining the capability to form tube-like structure. Results The isolated cells grew in a monolayer and exhibited mosaic-like pattern under an inverted phase-contrast microscope. They strongly expressed CD31, an endothelium-specific surface marker, and had an obvious capability to form tube-like structure. Conclusion The method established for isolation of HUAECs in this experiment is simple, rapid and efficient. In addition, the isolated cells are of high purity and high survival.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第7期759-762,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
北京大学985专项基金(985II/56210405)
关键词
人脐动脉
内皮细胞
分离
原代培养
细胞鉴定
human umbilical artery
endothelial cells
isolation
primary culture
cell identification
