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Silke Oellerich, Daniele Spinozzi, Alina Miron, Isabel Dapena, Mehrdad Rafat, Gerrit Melles; In vitro evaluation and transplantation of human corneal endothelial cells cultured on biocompatible carriers. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2183.
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© ARVO (1962-2015); The Authors (2016-present)
To compare the adaptability of cultivated human corneal endothelial cells (hCEC) on human anterior lens capsules (HALC), bioengineered collagen sheets and denuded Descemet membranes (dDM) and to evaluate in vitro the suitability of the cell-carrier sheets as tissue-engineered grafts for Descemet membrane endothelial keratoplasty (DMEK).
hCEC were isolated, cultured up to P2 and seeded onto HALC, LinkCell™ bioengineered matrices of 20µm thickness (LK20) and dDM. hCEC morphology, viability and proliferation were assessed by light microscopy, cell viability assay and immunohistochemistry (Ki67, ZO-1 and Na+/K+-ATPase expression). hCEC-carrier constructs were evaluated by simulating DMEK surgery in vitro using an artificial anterior chamber (AC) and a human donor cornea without DM and were compared to an in vitro surgery with a standard DMEK-graft.
hCEC cultured on all carriers formed a monolayer of tightly packed cells that retained their proliferative capacity with a high cell viability rate and expression of phenotypical markers ZO-1 and Na+/K+-ATPase. During in vitro surgery, hCEC-HALC constructs behaved similar to the DMEK reference model during implantation and unfolding in the artificial AC and showed good adhesion to the bare stroma. hCEC-LK20 and hCEC-dDM constructs required some additional handling because of challenges related to rolling of the grafts and loading them in the injector, but both constructs could be implanted and unfolded in the artificial AC using a DMEK-like technique. hCEC-dDM constructs showed similar graft adherence as hCEC-HALC constructs, while adhesion of hCEC-LK20 constructs was poor.
All analyzed carriers were suitable substrates for cultivating hCEC. Upon in vitro DMEK surgery, hCEC-HALC constructs behaved similarly to a standard DMEK-graft on all scored aspects, while graft adhesion and surgical handling, respectively, are issues that still need to be addressed for hCEC-LK20 and hCEC-dDM constructs.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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