Corneal endothelial cells derived from monkey iPS cells: a short term evaluation

Abstract

Purpose: To evaluate the short term function of tissue-engineered corneal endothelial cells (TECE cells) derived from monkey induced pluripotent stem cells (iPS cells).

Methods: Cynomolgus monkey iPS cells were cultured in KSR medium for 1 week and subsequently in N2 medium for 1 week, supplemented with TGF-beta inhibitor and BMP inhibitor. The iPS-derived neural crest cells were isolated as CD271 positive fraction by cell sorter. Next these cells were proliferated with EGF and FGF2, and subsequently medium was changed to an “endothelium-deriving medium” including GSK-3beta inhibitor, retinoic acid and ROCK inhibitor. These cells were dispersed on collagen sheet and TECE cell sheets were obtained. The pump function attributable to Na,K-ATPase activity of TECE cell sheets was measured with an Ussing chamber, and compared with that of human corneal endothelial cell line (B4G12 cells). In vivo function of TECE was measured as central corneal thickness of rabbit eyes transplanted with TECE cell sheets for 8 days after surgery and compared with control eyes deprived of endothelium.

Results: Hexagonal mosaic pattern monolayer TECE cells were obtained. Pump function of TECE was 2.34±0.46 mV, whereas that of B4G12 cells was 1.07±0.20 mV. The corneal thickness of TECE transplanted rabbit eyes (589.25±164.8μm) maintained significant lower corneal thickness than control eyes (1105.8±165.9μm) throughout the post-operative period.

Conclusions: In vitro and short term in vivo function of monkey iPS-derived TECE were observed. Further long-term in vivo evaluation of TECE transplantation to monkey eyes is needed.

Keywords: 481 cornea: endothelium • 721 stem cells • 606 NaK ATPase