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Invest Ophthalmol Vis Sci ; 60(15): 5095-5103, 2019 12 02.
Article in English | MEDLINE | ID: mdl-31826231

ABSTRACT

Purpose: To establish Myh11 as a marker of a subset of corneal endothelial cells (CECs), and to demonstrate the feasibility of restoring the corneal endothelium with Myh11-lineage (Myh11-Lin[+]) adipose-derived stromal cells (ASCs). Methods: Intraperitoneal administration of tamoxifen and (Z)-4-hydroxytamoxifen eyedrops were used to trace the lineage of Myh11-expressing cells with the Myh11-Cre-ERT2-flox-tdTomato mouse model. Immunostaining and Western blot characterized marker expression and spatial distribution of Myh11-Lin(+) cells in the cornea, and administration of 5-ethynyl-2'-deoxyuridine labeled proliferating cells. ASCs were isolated from epididymal adipose Myh11+ mural cells and treated with cornea differentiation media to evaluate corneal endothelial differentiation potential. Differentiated ASCs were injected into the anterior chamber to test for incorporation into corneal endothelium following scratch injury. Results: A subset of CECs express Myh11, a marker previously thought restricted to only mural cells. Myh11-Lin(+) CECs marked a stable subpopulation of cells in the cornea endothelium. Myh11-Lin(+) ASCs undergo CEC differentiation in vitro and incorporate into injured corneal endothelium. Conclusions: Dystrophy and dysfunction of the corneal endothelium accounts for almost half of all corneal transplants, the maintenance of the cornea endothelium is poorly understood, and there are a lack of mouse models to study specific CEC populations. We establish a mouse model that can trace the cell fate of a subpopulation of CECs based on Myh11 expression. A subset of ASCs that share this Myh11 transcriptional lineage are capable of differentiating into CECs that can incorporate into injured corneal endothelium, revealing a potential cell source for creating engineered transplant material.


Subject(s)
Adipocytes/transplantation , Corneal Dystrophies, Hereditary/metabolism , Corneal Transplantation/methods , Endothelium, Corneal/metabolism , Myosin Heavy Chains/metabolism , Stromal Cells/transplantation , Animals , Cell Count , Cell Differentiation , Cells, Cultured , Corneal Dystrophies, Hereditary/pathology , Corneal Dystrophies, Hereditary/surgery , Disease Models, Animal , Endothelium, Corneal/pathology , Immunoblotting , Immunohistochemistry , Mice
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