Long-term cultivation of human corneal endothelial cells by telomerase expression

被引：5

作者：

Liu, Zhiping ^{[1
]}

Zhuang, Ling ^{[1
]}

Li, Chaoyang ^{[1
]}

Wan, Pengxia ^{[1
]}

Li, Naiyang ^{[1
]}

Zhou, Qiang ^{[1
]}

Zhou, Chenjing ^{[1
]}

Huang, Zheqian ^{[1
]}

Wang, Zhichong ^{[1
]}

机构：

[1] Sun Yat Sen Univ, Zhongshan Ophthalm Ctr, State Key Lab Ophthalmol, Guangzhou 510060, Guangdong, Peoples R China

来源：

EXPERIMENTAL EYE RESEARCH | 2012年 / 100卷

基金：

中国国家自然科学基金; 国家高技术研究发展计划(863计划);

关键词：

cornea; corneal endothelial cells; cell line; telomerase; LARGE T-ANTIGEN; CONTACT-LENS WEAR; EPIDERMAL-GROWTH-FACTOR; EPITHELIAL-CELLS; REPLICATIVE SENESCENCE; PROLIFERATIVE CAPACITY; DESCEMETS-MEMBRANE; ALPHA-THALASSEMIA; TIGHT JUNCTION; HUMAN LIMBAL;

D O I：

10.1016/j.exer.2012.04.013

中图分类号：

R77 [眼科学];

学科分类号：

100212 ;

摘要：

The objective of this study was to explore the potential role of human telomerase reverse transcriptase (TERT) in extending the proliferative lifespan of human corneal endothelial cells (HCECs) under long-term cultivation. A primary culture was initiated with a pure population of HCECs in DMEM/F12 media containing 10% fetal bovine serum and other various supplements. TERT gene was successfully transfected into normal HCECs. A stable HCECs cell line (TERT-HCECs) that expressed TERT was established. The cells could be subcultured for 36 passages. Within this line of cells, TERT not only extended proliferative lifespan and inhibited apoptosis but also enhanced the cell line remaining the normal characteristics similar to HCECs. There were no significantly differences in the expression of the pump function related proteins voltage dependent anion channel 3 (VDAC3), sodium bicarbonate cotransporter member 4 (SLC4A4), chloride channel protein 3 (CLCN3), Na+/K+-ATPase alpha 1, and ZO-1 in the cell line TERT-HCECs and primary HCECs. TERT-HCECs formed a monolayer cell sheet, maintained similar cell junction formation and pump function with primary HCECs. Karyotype analysis exhibited normal chromosomal numbers. The soft agar colony assay and tumor formation in nude mice assay showed no malignant alterations in TERT-HCECs. Our findings indicated that we had established a cell line with its similar phenotype and properties to primary HCECs. Further study of the TERT-HCECs may be valuable in studying the function of the cells in vivo. (c) 2012 Elsevier Ltd. All rights reserved.

引用

页码：40 / 51

页数：12

共 85 条

[1] Morphological and functional analysis of immortalized human corneal endothelial cells after transplantation [J].