Extended Data Figure 2 : Limbal biopsies from normal donors or patients with LSCD.

From: ABCB5 is a limbal stem cell gene required for corneal development and repair

Extended Data Figure 2

ac, Limbal biopsies were obtained from normal donors or patients with LSCD. a, Typical findings are shown for a patient (patient 1) with a chemical burn before receiving a penetrating keratoplasty plus kerato-limbal allograft from a cadaveric donor eye (donor 1). Serial cross-sections of the biopsies were stained with either H&E, isotype control monoclonal antibody, or ABCB5 moncolonal antibody. ABCB5 staining in the limbal epithelium of donor 1 reveals nests of ABCB5-positive cells, whereas ABCB5-positivity is reduced in the limbal epithelium of patient 1. Photographs of immunofluorescent staining are montages of sequential photos at ×20 magnification. In these studies, equal-sized biopsies were recovered from a portion of the patient and donor limbus, frozen, and sectioned to produce eight sequential sections. All epithelial cells were counted in each section. A total of 2,031 and 2,051 epithelial cells were counted in patient 1 and donor 1, respectively. b, Limbal biopsies were obtained from a patient (patient 2) with an autoimmune corneal melt, peripheral ulcerative keratitis, and partial limbal stem cell deficiency before receiving a kerato-limbal autograft from the patient’s normal contralateral eye (donor 2). Serial sections of the biopsies were stained with either H&E, isotype control monoclonal antibody, or ABCB5 monoclonal antibody. ABCB5 positivity was present in the basal layer of the limbal epithelium of donor 2, while a dramatically reduced epithelial layer and no ABCB5 staining was observed in the limbus of patient 2. Photographs of immunofluorescent staining are montages of sequential photos at ×20 magnification. In these studies, equal-sized biopsies were recovered from a portion of the patient and donor limbus, frozen, and sectioned to produce eight sequential sections. All epithelial cells were counted in each section. A total of 563 and 2,662 epithelial cells were counted in patient 2 and donor 2, respectively. Patient 2 had a reduced number of epithelial cells due to the extensive damage from chronic autoimmunity. c, LSCD patient information. *Donor 1: cadaveric donor; **Donor 2: autologous transplant from contralateral eye. KLAL, kerato-limbal allograft (limbal tissue was harvested from donor eye); KLAU, kerato-limbal autograft (part of limbal tissue was resected from uninjured contralateral eye); OD, right eye; PKP, penetrating keratoplasty; PUK, peripheral ulcerative keratitis.