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Exp Eye Res. 2016 Oct;151:107-14. doi: 10.1016/j.exer.2016.08.004. Epub 2016 Aug 12.

Directed differentiation of human embryonic stem cells to corneal endothelial cell-like cells: A transcriptomic analysis.

Author information

1
Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China; Department of Human Genetics, David Geffen School of Medicine, UCLA, 695 Charles Young Drive South, Los Angeles, CA 90095, United States.
2
Department of Human Genetics, David Geffen School of Medicine, UCLA, 695 Charles Young Drive South, Los Angeles, CA 90095, United States.
3
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun-Ye-Sat University, Guangzhou, 510060, China.
4
Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. Electronic address: liuqh@njmu.edu.cn.
5
Department of Human Genetics, David Geffen School of Medicine, UCLA, 695 Charles Young Drive South, Los Angeles, CA 90095, United States; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun-Ye-Sat University, Guangzhou, 510060, China. Electronic address: gfan@mednet.ucla.edu.

Abstract

Corneal endothelial cells (CECs) are a monolayer of cells covering the inner-side of cornea, playing a pivotal role in keeping the cornea transparent. Because adult CECs have no proliferative capacity, the loss of CECs during aging or under pathological conditions would lead to corneal edema, eventually leading to the blindness. Clinically, donated CECs have been successfully transplanted to treat the diseases of CEC deficiency; however, the source of CEC donation is very limited. As an alternative cell source for CEC transplantation, CEC-like cells can be obtained via in vitro differentiation of human pluripotent stem cells. In this study, we introduced a modified two-stage differentiation method to convert H9 human embryonic stem cells (hESCs) to neural crest cells (NCCs), then further into CEC-like cells. The CEC-like cells treated with bovine CEC conditional medium morphologically best resembled primary CECs among all the culture conditions. By whole transcriptome analysis, we found that the typical markers of CECs, such as Na+-K+-ATPase, AQP1, Col8a and ZO-1, are highly expressed in hESC-derived CEC-like cells. By comparing RNA transcriptome of hESC-derived CEC-like cells with human primary fetal and adult CECs, we further identified shared molecular markers such as TRIT1, HSPB11, CRY1 that can be used to quality control CEC derivatives from hESCs. Our study paves the way for the quality-control and future application of hESC-derived CECs in the treatment of CEC deficiency disorders.

KEYWORDS:

Corneal endothelial cells; Directed differentiation; Human embryonic stem cells; Transcriptomic analysis

PMID:
27523468
DOI:
10.1016/j.exer.2016.08.004
[Indexed for MEDLINE]

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