Generation and purification of functional corneal endothelium-like cells differentiated from human embryonic stem cells

Abstract

Purpose : To restore endothelial function in patients with corneal endothelium dysfunction through cell transplant therapies, here we study differentiation of human embryonic stem cells (hESCs) into corneal endothelial cells (HCECs), and their properties of barrier and pump function.

Methods : A two-step method was used first to differentiate stem cells into neural crest cells (NCC), and then to induce maturation to HCEC-like cells. Cell morphology, phenotypic markers including expression of specific markers, and barrier functional assays assessed by transendothelial electrical resistance (TEER) were detected along a time course after differentiation.

Results : Differentiated hESC-derived HCECs demonstrated similar polygonal morphology as canonical primary HCECs. HCEC-specific markers, including zonula occludens-1 (ZO-1), sodium-potassium ATPase (Na⁺-K⁺-ATPase) and collagen VIII, showed similar expression patterns in hESC-derived HCEC-like cells. Cell sorting using different surface markers revealed that subpopulations with double-positive CD56 and CD166 marker expression showed higher barrier function by TEER than subpopulations positive for only a single HCEC marker.

Conclusions : Human stem cells can generate a large yield of HCEC-like progeny that carry phenotypic and functional measures similar to primary HCECs.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.