人脐带动脉与脐带静脉血管内皮细胞在体外相同培养环境下的定性研究  

Differential Characterization of Human Umbilical Artery and Umbilical Vein Endothelial Cells under Same Culture Conditions In Vitro

作　　者：刘倩 刘通海 叶进培 LIU Qian;LIU Tonghai;YE Jinpei(The Institute of Biomedical Sciences,Shanxi University,Taiyuan 030006,China)

机构地区：[1]山西大学生物医学研究院,太原030006

出　　处：《中国细胞生物学学报》2020年第4期656-663,共8页Chinese Journal of Cell Biology

摘　　要：血管内皮细胞受损与许多心脑血管疾病的发生和发展有关,已成为当今生命科学、药学领域的重要的研究对象。体外分离获得血管内皮细胞以及鉴定对于研究血管功能和为心脑血管疾病建立细胞模型极为重要。该研究通过了一种简便快捷、分离纯度高的人脐带动脉内皮细胞(human umbilical cord artery endothelial cells,HUAEC)和脐带静脉内皮细胞(human umbilical cord vein endothelial cells,HUVEC)的分离及培养体系,同时从形态特征、增殖活力、成管能力、表面抗原和特异基因的表达等方面检测了二者在体外相同培养环境下的动态变化与差异。该研究发现,HUAEC和HUVEC在体外连续传代培养过程中形态特征、成管能力、表面抗原(CD144、CD31、CD309、CD133、CD34)这几个方面作为内个皮细胞的基本特性没有明显差异,尽管HUAEC相对于HUVEC增殖活力更高。对于新鲜分离的HUAEC和HUVEC,二者特异性基因表达水平具有显著差异(HUAEC高表达EFNB2、DLL4、NRP1、CXCR4;HUVEC高表达EPHB4、COUP-TF II),然而随着培养时间的延长(传代次数的增加),HUAEC丧失其特异性基因(P6)的表达之后,HUVEC仍保持其特异性基因的高表达。因此HUVEC特异性表达基因EPHB4、COUP-TF II可以作为区分体外培养的人脐带动脉或者脐带静脉来源的内皮细胞的可靠鉴定标志基因。The damage of vascular endothelial cells is related to the development of many cardiovascular and cerebrovascular diseases,which has become an important research subject in the fields of life science and pharmaceutical industry.In vitro isolation and identification of vascular endothelial cells is important for studying vascular function and establishing a cellular model for cardiovascular and cerebrovascular diseases.The present study has used a simple and rapid isolation and culture system for HUAEC(human umbilical cord artery endothelial cells)and HUVEC(human umbilical cord vein endothelial cells)with a high purity,and investigates systematically the dynamic changes in biological characteristics including morphology,proliferation,tube formation,surface antigen and the expression of specific genes of both HUAEC and HUVEC under the same culture conditions in vitro.This research found that there was no significant difference in the morphology,tube forming ability,surface antigen(CD144,CD31,CD309,CD133,CD34)between HUAEC and HUVEC for a long-term continuous subculture,although HUAEC showed a higher proliferation activity than HUVEC.In freshly isolated HUAEC and HUVEC,the expression levels of the specific genes for artery and vein respectively were significantly different(HUAEC high expression in EFNB2,DLL4,NRP1,CXCR4;HUVEC high expression in EPHB4,COUP-TF II).However,during the prolonged culture period(after passage 6)HUAEC lost then its specific gene expression,whereas HUVEC remained its high expression of the specific genes.In conclusion,HUVEC-specific expression genes EPHB4 and COUP-TF II can be used as reliable identification markers for distinguishing human umbilical cord artery-from umbilical cord vein-derived endothelial cells cultured in vitro.

关 键 词：人脐带动脉内皮细胞 人脐带静脉内皮细胞 成管功能 表面抗原 特异性基因 内皮细胞定性 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]