Small-Molecule Induction Promotes Corneal Epithelial Cell Differentiation from Human Induced Pluripotent Stem Cells
Authors
Alexandra Mikhailova, Tanja Ilmarinen, Hannu Uusitalo, and Heli Skottman
Institution
Uni Tampere
Country
Finland
Year
2014
Journal
Stem Cell Reports
Abstract
Human induced pluripotent stem cells (hiPSCs) offer unique opportunities for developing novel cell-based therapies and
disease modeling. In this study, we developed a directed differentiation method for hiPSCs toward corneal epithelial progenitor
cells capable of terminal differentiation toward mature corneal epithelial-like cells. In order to improve the efficiency and reproducibility
of our method, we replicated signaling cues active during ocular surface ectoderm development with the help of two smallmolecule
inhibitors in combination with basic fibroblast growth factor (bFGF) in serum-free and feeder-free conditions. First, smallmolecule
induction downregulated the expression of pluripotency markers while upregulating several transcription factors essential
for normal eye development. Second, protein expression of the corneal epithelial progenitor marker p63 was greatly enhanced, with
up to 95% of cells being p63 positive after 5 weeks of differentiation. Third, corneal epithelial-like cells were obtained upon further
maturation.