大鼠股动脉血管原代内皮细胞的分离与培养  

Isolation and culture of primary endothelial cells from rat femoral artery

作　　者：刘倩 韩陈陈 罗婷婷 张雨 李浩 马旸[1] 魏伟[1] Liu Qian;Han Chenchen;Luo Tingting(Institute of Clinical Pharmacology,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学临床药理研究所,合肥230032

出　　处：《安徽医科大学学报》2021年第4期566-570,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81502123、81330081、81673444)。

摘　　要：目的体外分离、培养并鉴定大鼠股动脉血管原代内皮细胞,为相对较小血管来源的原代内皮细胞功能研究提供细胞模型。方法无菌取大鼠股动脉血管,胶原酶消化大鼠股动脉血管内皮细胞使其分离,流式细胞术检测内皮细胞纯度;流式分选出CD31阳性细胞,并用激光共聚焦鉴定内皮细胞;以CCK-8法、高内涵细胞成像法检测内皮细胞增殖能力,以Transwell法、Matrigel胶法检测其迁移和成管功能。结果胶原酶消化分离所得股动脉血管原代内皮细胞在14~16 d细胞覆盖培养瓶面积的2/3以上,细胞呈现典型铺路石状。流式检测其纯度为38.34%,分选后利用CD31进行荧光染色鉴定,细胞均为CD31阳性;PGE2可显著上调CD31阳性内皮细胞增殖、迁移、成管功能。结论该研究采用简便的方法成功分离得到股动脉血管原代内皮细胞,为研究相对较小血管来源的原代内皮细胞功能提供体外细胞模型。Objective To isolate,culture and identify rat femoral artery endothelial cells in vitro,and to provide an in vitro cell model for pathological and physiological studies.Methods The rat femoral artery blood vessels were taken aseptically,and rat femoral artery endothelial cells were isolated by collagenase digestion;the purity of endothelial cells was detected by flow cytometry;CD31 positive cells were sorted out by flow cytometry,and endothelial cells were identified by laser confocal;the proliferation ability of endothelial cells was detected by CCK-8 method and high-content cell imaging method,and their migration and tube function were detected by Transwell method and Matrigel gel method.Results The primary endothelial cells of the femoral artery vascular obtained by collagenase digestion and separation had covered more than 2/3 of the area of the culture flask at 14-16 d,and the cells were typical paving stones.The purity was 38.34%,and the sorted cells were identified by CD31 fluorescence staining.The cells were all CD31 positive;PGE2 can significantly upregulate the proliferation,migration,and tube formation of CD31 positive endothelial cells.Conclusion In this study,a simple method is used to successfully isolate the primary endothelial cells of femoral arteries,providing an in vitro cell model for studying the function of primary endothelial cells derived from relatively small blood vessels.

关 键 词：大鼠 股动脉 原代血管内皮细胞 分离培养 鉴定 

分 类 号：R394.26[医药卫生—医学遗传学] R322.121[医药卫生—基础医学]