Abstract

AbstractPurposeHuman corneal endothelial cells (HCECs) fail to proliferate sufficiently following pathological injuries presumably due to their cell cycle arrest in the G1 phase (Joyce, 2012; Joyce et al, 1996). The current treatment for corneal endothelial disease includes partial (endothelial keratoplasty) or completes (penetrating keratoplasty) transplantation of the cornea. The shortage of available donor corneas worldwide continues to limit transplantation and thus necessitates alternative compensatory approaches. Stimulating of HCECs in vivo to promote cell proliferation or bioengineering of corneal endothelium for transplantation may offer practical solutions. Several methods for in vitro expansion and maintenance of HCECs have been developed until date but it is difficult to judge their reproducibility. Additionally, the cellular and molecular biology of these cells are not fully understood. Therefore, a comprehensive gene expression study on requisite cell‐cycle processes and associated signaling cascades may help to establish an important step for engineering of a functional corneal endothelium.MethodsWe attempted to develop a three step culture system to understand the gene expression profiles as well as the associated signaling pathways of native, proliferated and bioengineered HCECs corneal endothelium at different steps by RNA sequencing (RNA seq), quantitative real time PCR (qRT‐PCR) and scanning electron microscopy (SEM).ResultsOur results characterized 80 genes as markers of HCECs throughout the cell‐cycle stages, primarily focused on the associated cell cycle signaling cascades. Engaged genes were mostly related to entire cell‐cycle stages including the regulators of cell cycle and the cell cycle related signaling pathways suggesting their association with cell proliferation.ConclusionThis study adds valuable information to the literature on biology of HCECs and forms a potential for prospective clinical applications of the data for advancing the field of corneal regenerative medicine.

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