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Biochem Biophys Res Commun. 2018 May 23;499(4):735-742. doi: 10.1016/j.bbrc.2018.03.168. Epub 2018 Apr 10.

Conditional reprogrammed human limbal epithelial cells represent a novel in vitro cell model for drug responses.

Author information

1
Shenzhen Eye Hospital, Shenzhen, Guangdong 518040, China; Shenzhen R&D Center of State Key Laboratory of Virology, Wuhan University Shenzhen Institute, Shenzhen, Guangdong 518057, China; State Key Laboratory of Virology, Institute of Medical Virology, Wuhan University School of Basic Medical Sciences, Wuhan, Hubei 430071, China.
2
Shenzhen Eye Hospital, Shenzhen, Guangdong 518040, China; Visual Optics Institute, Health Science Center, Shenzhen University, Shenzhen, Guangdong 518060, China.
3
Shenzhen Eye Hospital, Shenzhen, Guangdong 518040, China; Shenzhen R&D Center of State Key Laboratory of Virology, Wuhan University Shenzhen Institute, Shenzhen, Guangdong 518057, China.
4
Shenzhen R&D Center of State Key Laboratory of Virology, Wuhan University Shenzhen Institute, Shenzhen, Guangdong 518057, China.
5
Shenzhen Eye Hospital, Shenzhen, Guangdong 518040, China.
6
Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, China.
7
Shenzhen R&D Center of State Key Laboratory of Virology, Wuhan University Shenzhen Institute, Shenzhen, Guangdong 518057, China; State Key Laboratory of Virology, Institute of Medical Virology, Wuhan University School of Basic Medical Sciences, Wuhan, Hubei 430071, China. Electronic address: somhli@whu.edu.cn.

Abstract

In this study, we established human limbal epithelial cells (LECs) from normal limbal tissues by using Conditional Reprogramming (CR) technology (refer to CR-LEC cells in this study). We have successfully established CR-LEC cell strains from three human donors (3 out of 3), and normal rabbits (2 out of 2) and pig (1 out of 1) as well. CR-LEC cells sustained a continuous and stable proliferation status with a normal karyotype, normal response to DNA damage, well-defined structured spheres in matrigel 3D culture. Responses of CR-LEC cells to IFN α2b, Ganciclovir and 5-Fluorouracil were different, suggesting that these drugs had different toxicities to these cells as expected. More important, there was no significant difference of responses to drugs between early and late passages of CR-LEC cells (p>0.05), indicating CR-LEC cells can serve a stable normal human cell model for toxicity assessment. Toxicity tests with monolayer cultures of CR-LEC cells were measured by staining the F-actin and Dsg-1 expression. Toxicity of three drugs at LD50 concentration resulted in a gradually increased destruction of monolayer, which is, in accordance with the irritation grade of three drugs on human cornea epithelium. Therefore, CR-LEC cells provide a novel and reliable in vitro physiological cell model for corneal toxicity assessment.

KEYWORDS:

Cell viability; Conditional reprogramming technology; Drug toxicity assessment; Human limbal epithelial cell; Monolayer assay

PMID:
29577905
DOI:
10.1016/j.bbrc.2018.03.168
[Indexed for MEDLINE]

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