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Emi Inagaki, Shin Hatou, Kazunari Higa, Hideyuki Miyashita, Satoru Yoshida, Kazuo Tsubota, Shigeto Shimmura; Functional analysis of tissue engineered corneal endothelium from human skin derived precursors.. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3450.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal endothelial dysfunction remains a major indication for corneal transplantation. However, corneal transplantation has problems such as shortage of donors and graft rejection. Skin-derived precursors (SKPs) are post-natal stem cells of neural crest origin, similar to the corneal endothelium. In this study, we show the functional analysis of tissue engineered corneal endothelium derived from human SKPs.<br />
Human SKPs (hSKPs) were isolated from the eyelid skin taken from patients undergoing eyelid surgery and then cultured in SKPs proliferation medium. hSKPs-TECE (tissue engineered corneal endothelium) was induced in a medium containing retinoic acid and glycogen synthase kinase (GSK) 3b inhibitor (activator of Wnt/b-catenin signaling) by the method we previously reported using murine SKPs. To assess the function of TECE in vitro, we used the Ussing[S1] chamber which is an apparatus for measuring trans-endothelial resistance (TER) and pump function of monolayer cells. Human corneal endothelial cell line (B4G12 cell) was used as control.<br /> <br /> <br />
The TER of hSKPs-TECE was 17.96±4.16 (Ω/cm2), control B4G12 was 8.86±0.67 (Ω/cm2). Na,K-ATPase pump activity of SKPs-TECE was calculated by the difference of short-circuit current before and after Ouabine addition. Na,K-ATPase pump activity of SKPs-TECE was 2.64±0.32 (mV/cm2), control B4G12 was 0.79±0.03 (mV/cm2). hSKPs-TECE has significantly higher pump function (3.3-fold) compared to control B4G12 cells.<br />
We successfully induced functionally differentiated tissue engineered corneal endothelium from human postnatal stem cells derived from skin. This may lead to a novel autologous stem cell therapy for corneal endothelial function.<br />
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