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Xiaoniao Chen, Lingling Wu, Yifei Huang, Liqiang Wang; Directed differentiation of human corneal endothelial cells from human embryonic stem cells by using cell-conditioned culture media. Invest. Ophthalmol. Vis. Sci. 2018;59(9):2248.
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Shortage of human donor cornea for transplant may not be able to meet the clinical demand. It is especially true for the endothelial transplant due to the poor proliferative nature of endothelia. Stem cell-based therapy is considered as one of the promising methods to solve the above problems. However, the induced differentiation of embryonic stem cells (hESCs) is complicated and not feasible for massive production, therefore we developed a simple and straightforward approach for endothelial cell differentiation from hESCs.
Human embryonic stem cells are used to differentiate into human corneal endothelial cells by using a two-stage method, in which involving the application of two different types of conditioned culture medium. The simulation of the developmental environment of embryonic tissue in the anterior ocular segment is achieved by using the conditioned media derived from human corneal stromal cells (HCSC) and human lens epithelial cells (LECs) cultures.
In the presence of conditioned culture medium, embryonic stem cells were differentiated first towards the fate of periocular mesenchymal precursors (POMPs) in the context of expression of several POMP markers (AP2α, p75, CD57). Following this first stage differentiation, POMPs were further directed to differentiate into corneal endothelia cell (CEC)-like cells in the presence of second conditioned culture medium. The differentiation of POMPs into CEC-like cells is regulated by TGFβ2/FOXC1 signaling pathway that is activated by the factors presenting in the conditioned culture medium. According to the positive results of specific markers in immunofluorescent microscopy, we could initially identify the directed differentiation of target cells from hESCs.
Human corneal endothelia cell (HCEC)-like cells could be differentiated from human embryonic stem cells by simply using a 2-step pre-conditioned medium mediated approach, we presented a novel protocols for the generation of CEC-like cells, which are phenotypic and genetic similar to normal human CECs from hESCs. The results of this study may provide supportive evidence for the embryonic development of the cornea; also, it could serve as a potential strategy for corneal regeneration.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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