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J. Wang, J. Liu, D.W. Li; Establishment of Human Lens Epithelial Cell Line Expressing the Human Telomerase Gene . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1224.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Our previous studies have revealed that the human telomerase catalytic subunit (hTERT) has a variety of functions when introduced into different lines of lens epithelial cells (Xiang et al., 2000, BBRC, 278: 503-510; 2002, Oncogene, 21:3784-3791). To further study the function of hTERT in lens system, We have established human lens epithelial cell line expressing hTERT. Methods: Primary cultures of human lens epithelial cells isolated from patient donors were transfected with an exogenous human telomerase gene, which is under regulation of a CMV promoter. Individual stable expression clones were screened with G418. The growth rate and other properties of the hTERT-transfected cells were examined with various techniques. Results:RT-PCR confirmed that the exogenous hTERT gene is expressed in human lens epithelial cells. TRAP assays revealed that the exogenous hTERT maintains the telomerase activity of the transfectged human lens epithelial cells. The hTERT-transfected stable clones have been passaged for more than 40 population doubling generations, displaying normal growth pattern. In contrast, the parental primary cultures and the vector-transfected primary cultures have no telomerase activity. The parental primary cultures could not grow in G418-containing media and grow for a limited population doubling generations in G418-free MEM. The vector-transfected cells quickly run into senescence Conclusions:The exogenous human telomerase gene maintain lens epithelial cell growth and prevent senescence of the transfected human lens epithelial cells. The established human lens epithelial cell line will provide a useful model for other studies of lens biology and pathology. Supported by EY 11372 and the Hormel Foundation. None.
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