Small-molecule compounds promote corneal epithelial cell differentiation from human embryonic stem cells

  Background  Limbal stem cell deficiency is the main cause of corneal blindness, and the limited source of seed cells is the bottleneck of cell transplantation therapy.

  Objective  To induce human embryonic stem cells (hESCs) to differentiate into corneal epithelial progenitor cells and mature corneal epithelial cells.

  Methods  The induction of human embryonic stem cell line H9 (hESCs H9) involved embryonic body (EB) formation and the addition of small molecular compounds (SB505124 and IWP-2) and basic fibroblast growth factor (bFGF). After observation of cell morphology, the cells were collected at day 10, 15 and 20 of induction, respectively, and the mRNA and protein expression of characteristic markers OCT4, SOX2, ΔNp63; CK14, CK3, CK12 were detected by immunofluorescence assay and quantitative real-time PCR.

  Results  The morphology of induced cells was different from that of hESCs and similar to that of corneal epithelial cells (CECs). Compared with hESCs, the expressions of stem cell markers OCT4 and SOX2 were down-regulated (P＜0.01, respectively), and the expressions of corneal epithelial progenitor cells and corneal epithelial cell markers ΔNp63, CK14, CK3, CK12 were up-regulated after induction (all P＜0.05).

  Conclusion  This small-molecule compound promotes corneal epithelial cell differentiation from human embryonic stem cells.