精准医学杂志2024，Vol.39Issue(2)：134-138,143,6.DOI:10.13362/j.jpmed.202402008
小鼠角膜上皮细胞永生化细胞系的建立及表型鉴定
Establishment and phenotypic identification of an immortalized mouse corneal epithelial cell line
摘要
Abstract
Objective To investigate the method for establishing an immortalized mouse corneal epithelial cell line,and to identify the stability of its phenotype.Methods Primary mouse corneal epithelial cells were extracted and transfected with Simian Vacuolating Virus 40 Large T Antigen(SV40-LT)lentivirus to establish an immortalized cell line.HE staining and immu-nofluorescent staining were used to identify the phenotypes and markers(K12,Pax6,Ki-67)of corneal epithelium.Primary cor-neal epithelial cells were extracted to identify K14 protein in corneal epithelial cell layer,and bright field observation was performed to evaluate cell morphological characteristics.Several markers(K12,Pax6,Ki-67,ABCG2,K14)were identified for P1 and P8 cells after transfection.Crystal violet staining was used to observe the colony-formation ability of P1 and P8 cells after transfection.Results Mouse corneal epithelium was stained positive for K12,Pax6 and Ki-67,while the P1 and P8 generations of mouse cor-neal epithelial cells were stained positive for K12,Pax6,Ki-67,K14,and ABCG2;there were no significant differences in the posi-tive rates of the above markers(P＞0.05).There was no significant difference in colony-formation ability between P1 and P8 cells(P＞0.05).Conclusion The immortalized mouse corneal epithelial cell line is successfully established by transfection with SV40-LT lentivirus,which has stable phenotype and proliferation.关键词
细胞系/上皮细胞/角膜/转染/遗传标记/小鼠,近交C57BLKey words
Cell line/Epithelial cells/Cornea/Transfection/Genetic markers/Mice,inbred C57BL分类
医药卫生引用本文复制引用
白莹,王一卉,葛欢欢,卞雯晗,陈鹏..小鼠角膜上皮细胞永生化细胞系的建立及表型鉴定[J].精准医学杂志,2024,39(2):134-138,143,6.基金项目
国家自然科学基金资助项目(81970782) （81970782）
