Abstract
Purpose :
To survey the effects of fortified post-thaw human amniotic membranes (HAMs) by platelet lysate (PL) on proliferation and maintenance of overlying cultivated human limbal stem cells (hLSCs).
Methods :
PL was obtained from 3 volunteers by adding thrombin solution in 1:5 ratio. HAMs were frozen and thawed followed by fortifying with PL. hLSCs derived from donated corneoscleral rings were cultured on post-thaw fortified HAMs and compared with those on post-thaw non-fortified HAMs (controls) in terms of cell viability and genotypic / phenotypic alterations.
Results :
Cultivated hLSCs on post-thaw fortified HAMs, as compared to the controls, demonstrated a significant increase of viability on days 1 (P = 0.029) and 3 (P = 0.042), and a borderline increase after 1 week (P = 0.069). Gene expression profile showed a remarkable reduction of Tp63 on day 7 (P = 0.028) and a significant increase of NGFR on days 1, 3, and 7 (P = 0.028, P = 0.0001, and P = 0.0001, in that order). However, the expression of p63 protein was not significantly different between the cultivated LSCs on post-thaw fortified HAMs and the controls after 1 week (P = 0.18).
Conclusions :
Post-thaw HAMs fortified by PL have a favorable effect on proliferation and differentiation of cultured hLSCs from stem cell identity to epithelial precursor stage, and can be advantageous over non-fortified HAMs for ocular surface wound healings and reepithelialization.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.