The establishment of primary and transformed human vascular endothelial cell models
10.16352/j.issn.1001-6325.2025.12.1600
- VernacularTitle:原代及转化的人血管内皮细胞模型的建立
- Author:
Hailiang FENG
1
;
Linghua KONG
;
Jiayin DAI
;
Zhenli YANG
;
Xiaocui BIAN
;
Yuqin LIU
Author Information
1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 细胞资源中心,北京 100005
- Keywords:
vascular endothelial cells;
human umbilical vein endothelial cells;
SV40;
LT antigen;
CD31;
Cas9
- From:
Basic & Clinical Medicine
2025;45(12):1600-1607
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish primary and simian virus 40(SV40)T antigen transformed human vascular en-dothelial cell models,and provide available resources for endothelial research.Methods Human umbilical vein endothelial cells(HUVEC),human umbilical artery endothelial cells(HUAEC),great saphenous vein endothelial cells(GSVEC)and endothelial cells form endometrium and liver tissue were isolated and cultured respectively.Then,the primary endothelial cells were transformed by lentivirus containing SV40 big T and small T antigens,and continuously subcultured in vitro.The expression of CD31 was detected by flow cytometry,species identification-and mycoplasma detection by PCR,and cell identity was identified by STR detection.The transformed ECs were checked for HLA types.Some of them were tested for RNA expression profile and infected by Cas9 lentivirus to es-tablish stable clones.Results Totally 187 cell lines of transformed HUVEC,1 of transformed HUAEC,5 of trans-formed GSVEC,1 of transformed endothelial cells from endometrium and 1 of transformed endothelial cells from liv-er tissue,and 9 monoclonal HUVEC cell lines stably expressing Cas9 protein were established.All the transformed umbilical endothelial cells were CD31 positive ranging from 20%-90%for 20 cases,while for the rest 168 cases the positive rate was more than 90%.RNA expression revealed stable activation of cell proliferation(cell cycle and DNA synthesis).Their species were identified as human origin.The STR results were consistent with those of the primary culture and unique,and there was no mycoplasma contamination.All these cells could be obtained with the sharing services of National Science and Technology Infrastructure,the National Biomedical Cell-line Resource cen-ters(NSTI-BMCR).Conclusions A series of primary and SV40 T antigen transformed human vascular endothelial cell models have been established,which provide a tool for the study of cardiovascular diseases,inflammation,tumors and immune-related diseases.
