基础医学与临床2025，Vol.45Issue(12)：1600-1607,8.DOI:10.16352/j.issn.1001-6325.2025.12.1600
原代及转化的人血管内皮细胞模型的建立
The establishment of primary and transformed human vascular endothelial cell models
摘要
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.关键词
血管内皮细胞/人脐静脉内皮细胞/SV40/大T抗原/CD31/Cas9Key words
vascular endothelial cells/human umbilical vein endothelial cells/SV40/LT antigen/CD31/Cas9分类
生物科学引用本文复制引用
冯海凉,孔令华,代伽茵,杨振丽,卞晓翠,刘玉琴..原代及转化的人血管内皮细胞模型的建立[J].基础医学与临床,2025,45(12):1600-1607,8.基金项目
中国医学科学院医学与健康科技创新工程-重大协同创新项目(2021-I2M-1-053) （2021-I2M-1-053）
