Abstract
Purpose :
Corneal endothelial cells (CEnCs) are unable to proliferate in vivo and in the case of trauma or dystrophy the only treatment modality currently available is corneal or CEnCs transplant from cadaveric donor with high global shortage. Our ultimate goal is to 3D-bioprint the full-thickness human cornea using human stem cells. Here, we explored the possibility to bioprint human pluripotent stem cell (hPSC) derived CEnCs.
Methods :
Differentiated CEnCs from hPSCs were bioprinted using optimized hydrazone-crosslinked hyaluronic acid based bioink. Before bioprinting, the biocompatibility of the bioink with hPSC-CEnCs was first analyzed in vitro and further tested with transplantation on ex vivo denuded young porcine corneas. STEM121 was used to identify human origin cells from the ex vivo experiment. The viability of bioprinted hPSC-CEnCs was verified with live/dead stainings. Histological and immunofluorescence stainings with Zonula Occludens-1 (ZO-1), Sodium-potassium (Na+/K+)-ATPase and CD166 were used to confirm CEnC-like protein expression and morphology in all experiments.
Results :
The bioink modified for hPSC-CEnCs successfully supported the viability and printability of the cells. After 10 days of ex vivo transplantations, STEM121 positive hPSC-CEnCs were verified on the Descemet membrane of ex vivo porcine cornea showing the biocompatibility of the bioink. Seven days after bioprinting, the hPSC-CEnCs were viable and showed polygonal morphology with expression and native-like localization of ZO-1, Na+/K+-ATPase and CD166. Nevertheless, in some cases part of the cells underwent endothelial-mesenchymal transition and those cells spread underneath the CEnCs.
Conclusions :
Our results demonstrates that hPSC-CEnC can be bioprinted in covalently crosslinked hyaluronic acid bioink. This approach has potential as a corneal endothelium transplant and furthermore, can be used in the mission of the bioprinting the full-thickness cornea.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.