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Bioengineering of endothelial grafts using femtosecond Laser cut corneal lamellae or collagen lenticules, endothelialized with immortalized or hIPSC‐derived endothelial cells
Author(s) -
FOREST F,
HE Z,
MOULARD A,
SUFFEE N,
FOREST F,
DUMOLLARD JM,
PEOC'H M,
GAIN P,
THURET G
Publication year - 2014
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2014.4775.x
Subject(s) - decellularization , stromal cell , cornea , staining , chemistry , alizarin red , trephine , biomedical engineering , materials science , pathology , tissue engineering , anatomy , ophthalmology , biology , medicine
Purpose The bioengineering of endothelial grafts appears as a realistic solution to reduce the global scarcity for corneal donation worldwide. We focused our strategies of endothelial bioengineering on the endothelialization of human corneal stromal lamellae with hiPSC‐derived endothelial cells. Methods Human corneas unsuitable for graft but with a normal stroma, were cut into 4 to 7 lamellae of <100 μm thickness with a femtosecond laser (FsL), decellularized from their keratocytes using an optimized protocol (Ethanol/DNAse/SDS) and stored at +4°C. They were compared with collagen I compressed thin discs (RAFT) already described as carrier. Endothelialization was performed with immortalized endothelial cells (ECs) (BAG12) and hIPSC‐derived ECs (also presented during this congress). Efficiency was assessed by determining endothelial cell density (ECD), morphology after alizarin red staining and SEM and immunolabeling for Na/K‐ATPase expression. Transparency was measured using our customized transparometer Results both carriers were evenly endothelialized by B4G12 with similar alizarin red staining and Na/K ATPase expression patterns but higher ECD for stromal lamellae. Transparency was better for stromal lamellae. hIPSC‐derived ECs gave similar characteristics Conclusion Decellularized Fsl cut stromal lamellae seem better than collagen I compressed lenticules for the endothelial bioengineering using hISPC‐derived ECs. Unsuitable human cornea could be therefore “recycled” to increase the pool of available grafts. Grants: ABM2013, AIRE2013