Functional human corneal endothelial cell sheets harvested from temperature-responsive culture surfaces

SPECIFIC AIMOur aim was to fabricate self-supporting, bioengineered human corneal endothelial cell (HCEC) sheets in vitro that retain native morphological and functional characteristics, and thus are suitable for corneal regenerative therapies.PRINCIPAL FINDINGS1. DNA synthesis of HCECs on temperature-responsive culture dishesFor noninvasive cell sheet harvest that preserves cell viability, cell-to-cell junctions, extracellular matrix (ECM), and cell sheet contiguity, we applied unique temperature-responsive culture surfaces onto which an ultrathin layer of temperature-responsive polymer, poly (N-isopropylacrylamide) (PIPAAm), is covalently grafted. Since PIPAAm is known to undergo a phase transition across 32°C, PIPAAm-grafted culture surfaces are slightly hydrophobic (similar to ordinary tissue culture polystyrene) and facilitate cell adhesion under typical culture conditions at 37°C. Because of the spontaneous and rapid hydration of the grafted PIPAAm, adherent cultured cells release spontaneously from...