IHC characterization of 10- to 15-week-old adherent MCs

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IHC characterization of 10- to 15-week-old adherent MCs IHC characterization of 10- to 15-week-old adherent MCs. (A) H&E-stained sections of 10-week-old MC3 shows a thick stroma lined by a thin monolayer of epithelium-like and endothelium-like cells on either side and a lid margin (LM) on the top (arrowhead), connected by a thin periderm-like (P) continuous epithelial lining covering the entire ocular surface, with a fluid-filled lumen (L) in between. IHC characterization of 10- to 15-week-old adherent MCs. (A) H&E-stained sections of 10-week-old MC3 shows a thick stroma lined by a thin monolayer of epithelium-like and endothelium-like cells on either side and a lid margin (LM) on the top (arrowhead), connected by a thin periderm-like (P) continuous epithelial lining covering the entire ocular surface, with a fluid-filled lumen (L) in between. (B) Confocal images of tissue sections of MC3, immunostained for P63 (i), PAX6 (ii), VIM (iii), αSMA (iv), K12 and CD200 (v) (green) and counterstained with PI (red). Arrows point to the endothelial cell layer. (C) The corneal surface epithelium (arrow) of MC3 is a VIM− (ii), ciliary margin-like structure formed by ruffled pigmented epithelial cells (iv) and flanked by VIM+ ocular adnexal structures (iii). (D) H&E-stained sections of 15-week-old MC4 reveal mature cornea-like features, such as a thick stromal layer lined by a stratified squamous epithelium on the apical surface. A limbus-like structure separates the cornea-like epithelium and the goblet cell-enriched future conjunctiva. (E) Confocal images of tissue sections of MC4 immunostained with PAX6 antibody in red (i), or stained for P63 (ii), K12 (iii), VIM (iv), MUC2 (v) or αSMA (vii) in green and counterstained with PI in red. Brightfield IHC images are shown of sections stained with MUC5AC antibody (arrows) and Alcian Blue and counterstained with Hematoxylin or Nuclear Fast Red, respectively (vi). Praveen Joseph Susaimanickam et al. Development 2017;144:2338-2351 © 2017. Published by The Company of Biologists Ltd