Poster #017
Corneal epithelial progenitor cell differentiation from limbal-derived iPSC
Mentors: Vanessa Borges, PhD and Alexander Ljubimov, PhD
Limbal stem cell deficiency (LSCD) can cause discomfort, visual impairment, and blindness. A promising treatment approach involves using autologous induced pluripotent stem cells (iPSC). These iPSCs can be differentiated in vitro into limbal epithelial stem-like cells (LESC) and then transplanted back into patients. Our aim is testing the cryopreservation of pre-generated limbal-derived iPSCs and evaluating LESC differentiation. iPSCs were expanded and cryopreserved in culture medium supplemented with 10% DMSO or a commercial cell-freezing reagent, CryoStor®CS10. One week later, the cells were thawed and cultured again. Preservation in CryoStor®CS10 resulted in a significantly higher number of iPSC colonies, which were subsequently expanded and stored in CryoStor®CS10. Using these cells, we tested a previously established LESC differentiation protocol. Through brightfield and fluorescence microscopy, we observed that the cells differentiated into epithelial/limbal lineage, expressing several corneal markers, Pax6, ΔNp63, and some keratins. In conclusion, CryoStor®CS10-preserved limbal-derived iPSCs can successfully differentiate into limbal-like cells.