Isolation, cultivation and characterization of primary HCECs. A) Compact clusters of HCECs were released from the DM following approximately 5 hours of collagenase (2 mg/ml) treatment. B) A high magnification micrograph of dislodged HCECs sheet rounding up at the edges. C) Attachment of HCEC onto FNC-coated dish in a dispersed-clusters manner at Day 1 in the stabilization medium. D) Outspread and migration of HCECs in the proliferative medium from the second medium change onwards. E &F) Confluent HCECs displayed distinct cell borders and uniform cell shape in stabilization medium at Day 14. Confluent cultures of HCECs at the second passage express cellular markers indicative of the human corneal endothelium: G) Na+K+ATPase and native isotype-matched control (insert); as well as H) ZO-1 together with its native isotype-matched control (insert).