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Hun Lee, Ye Eun Han, Jimin Park, Sol Ah Han, Su Young Moon, Jae Yong Kim, Hungwon Tchah; Efficacy and safety of induced pluripotent stem cell-derived corneal endothelial-like cell transplantation in rabbit animal model of corneal endothelial failure. Invest. Ophthalmol. Vis. Sci. 2021;62(8):871.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the efficacy and safety of transplantation of the Peripheral blood mononuclear cell (PBMC) induced pluripotent stem cell (iPSC)-derived corneal endothelial-like cells into corneal endothelial failure animal model
PBMC iPSCs were generated using the iPS Reprogramming Kit with StemPro-34 medium and cultured under standard iPSC culture condition. Differentiation of iPSCs into corneal endothelial-like cells via neural crest cells (NCC) was induced using a conditioned medium in vitro. After 10 days culture of NCC with human endothelium serum free media, the phenotype of iPSC-derived corneal endothelial–like cells was detected by immunofluorescence. Related cell markers of iPSC-derived corneal endothelial-like cells were analyzed by quantitative real-time PCR. The cultured iPSC-derived corneal endothelial-like cells were transplanted into the rabbit anterior chamber by direct cell injection.
After 10 days culture of NCC with human endothelium serum free media, corneal endothelical cell-related markers, including zonula occludens-1 (ZO-1) and Na+/K+ ATPase, were expressed in the iPSC-derived corneal endothelial-like cells, showing well preserved hexagonality. The expression of ATP1A1, COL8A1, and AQP1 was higher in the iPSC-derived corneal endothelial-like cells, compared with the NCC and PBMC iPSCs. In animal experiment, increased corneal transparency was achieved after anterior chamber injection of the iPSC-derived corneal endothelial-like cells. PCR from the genomic DNA extract of central cornea which was enuclated 3 weeks after cell injection showed a positive result for human mitochondrial band, whereas negative band noted in the non-treated rabbit.
This preclinical study confirmed the therapeutic ability of the iPSC-derived corneal endothelial-like cells in vivo. Our findings demonstrated that iPSC-derived corneal endothelial-like cells could be a promising source for cell therapy in corneal endothelial dysfunction.
This is a 2021 ARVO Annual Meeting abstract.
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