Cadaveric human donor whole eye globes and corneas derived from consented donors according to Institutional Review Board (IRB)-approved protocols were obtained from the Saving Sight (Kansas City, MO) and CorneaGen (Seattle, WA) eye banks. The donor characteristics are described in Tables S1 and S2 . Limbal and central corneal epithelial cells were harvested as reported previously (Sasamoto et al., 2020 (link)). Briefly, the whole corneas were dissected from the eye globes and the central corneas were separated from the limbus using an 8 mm disposable biopsy punch (Integra LifeSciences, Plainsboro, NJ) and the corneal endothelium was removed mechanically. Limbal and central corneal epithelial cells were collected after 1-h incubation with PluriSTEM Dispase II Solution (MilliporeSigma, Burlington, MA) at 37°C and dissociated using TrypLE Express Enzyme (Thermo Fisher Scientific, Waltham, MA) at 37°C for 30 min. Cell cultures were maintained in DMEM/F12 medium (Thermo Fisher Scientific) supplemented with 10 ng/mL keratinocyte growth factor (KGF) (PeproTech, Rocky Hill, NJ), 10 μM Y-27632 (Tocris Bioscience, Bristol, UK) and B-27 Supplement (Thermo Fisher Scientific) (Miyashita et al., 2013 (link)).
293T cells (Clontech Laboratories, Mountain View, CA) were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific).
293T cells (Clontech Laboratories, Mountain View, CA) were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific).