Function Of BMP/Wnt Signaling In Regulation Of Limbal Stem Cells

Purpose: BMP and Wnt signaling pathways play an important role in regulation of limbal epithelial stem/progenitor cells,we carried out this study to explore how they function.Methods: Limbal segments were digested with Dispase II,single cells derived from limbal clusters were seeded with 3D-Matrigel(diluted in MESCM).We then activated or inhibited BMP and Wnt signaling separately and detected the level of p63??CK12?CEBP? and CK15 in LEPCs through q RT-PCR.We also detected the impression of Smad1/5/8 and?-catenin in LEPCs by immunofluorescence.The Wnt-activated LEPCs were used to treat LSCD rat model,immunofluorescence and H&E stain were applied to explore the repairing of limbus.Results: LEPCs aggregate and form spheres in 3D-Matrigel.BMP signaling up-regulate the impression of CEBP? and inhibit the impression of Ki-67.On the contrary,Wnt signaling up-regulate the impression of Ki-67.The blocking of BMP and Wnt signaling pathway had no significant effect on LEPCs stemness.The results of gel electrophoresis are consistent with that of real time.Wnt-activated LEPCs promote the regeneration of corneal epithelium and reduce the degree of corneal opacity in the treatment of LSCD rat model.Conclusions: Activation of BMP signaling pathway promotes LEPCs to remain quiescent state,Wnt signaling enhances LEPCs proliferation ability and leaves LEPCs in an undifferentiated self-renewal state.Wnt-activated LEPCs can promote the regeneration of corneal epithelium and reduce the degree of corneal opacity in the treatment of LSCD.The regulation of Wnt signaling pathway is expected to provide new ideas for the treatment of LSCD.