Fig. 1

A: a Limbus digested after 2-week plate seeding: f-LSC growth in monolayer (P0 culture passage, 10×); b example of an f-LSC limbosphere in low-adhesion culture conditions (20×); c, d example of an f-LSC limbosphere under adhesion culture condition (c 20×; d 40×). B: SSEA4⁺ immunofluorescence staining: a DAPI on a limbosphere; b SSEA4 detection on a limbosphere (20×); f DAPI on monolayer; g SSEA4 detection in f-LSC monolayer (40×); c, h merge; d, i DAPI/SSEA4-stained BM-MSCs; e, l DAPI/SSEA4-stained HeLa cells. C: Cytofluorimetric assay in total population (presorting, upper panel) and in SSEA4⁺ f-LSCs (postsorting, lower panel). Cells are negative for CD34, CD45, and HLA-DR. All fields are representative of one limbus sample out of at least 12 independent experiments. DAPI 4′,6-diamidino-2-phenylindole, N.C. negative control, P.C. positive control