Extended Data Figure 9: Corneal epithelial regeneration by transplantation in a rabbit LSC deficiency model.

a, Time course of corneal epithelial regeneration and repair in a rabbit LSC deficiency model post transplantation with GFP-labelled PAX6⁺ SESCs. Top panels, 3 day post transplantation. left, light micrograph showing a hazy cornea ; right, GFP+ donor cells at limbal region (arrows). Bottom panels, 20 days post transplantation. Left, light micrograph showing a cornea with partial clarity; right, GFP⁺ donor co-located in transparent areas (arrows). Scale bars, 1 mm. We observed that only the transplanted cells from the limbal region could survive, proliferate and regenerate cornea surface epithelium, suggesting that limbus contained the stem cell niche favourable for stem cell survival and growth. b, Culture and re-isolation of reprogrammed donor GFP-labelled PAX6⁺ SESCs epithelial cells from the limbal region of a rabbit recipient eye 90 days post transplantation with GFP-labelled PAX6⁺ SESCs. Top panel, double immunofluorescence staining of PAX6 and GFP; bottom panel, double immunofluorescence staining of p63 and GFP in PAX6-transduced rabbit SESCs. Scale bars, 100 µm. c, Repair and recovery of a repeat cornea epithelium injury on a cornea transplanted with GFP-labelled PAX6⁺ SESCs. Top panels, we iatrogenically scraped and removed donor-derived corneal epithelial cells and made a large corneal surface epithelium defect (arrows) 3 months post initial transplantation of PAX6⁺ SESCs. Bottom panels, complete repair and recovery were observed within 72 h with healed epithelial defect (n = 3). Left panels, light micrographs; middle panels, slit-lamp micrographs; right panels, fluorescein staining.