Effects Of Mechanical Properties Of Substrate Materials On DNA Methylation And Pluripotency Of Embryonic Stem Cells

Embryonic stem cells(ESCs)are pluripotency,self-renewing cells.More and more attention has been paid to the effect of the properties of extracellular matrix on the fate of stem cells.The elasticity and curvature of the substrate material have great influence on embryonic stem cell culture in vitro.DNA methylation is a major mechanism regulating gene expression.It is clear that strict regulation of DNA methylation is necessary for normal development.To explore the factors influencing embryonic stem cell culture in vitro,methylidene diacrylamide gel and polydimethylsiloxane(PDMS)were used as the substrate.Different elastic base materials were prepared by using methyldiacrylamide gel,and it was found that the soft base could maintain the DNA methylation level of embryonic stem cells at a low level,while the hard base significantly improved the DNA methylation level.This may be related to the cytofilament skeleton and adhesion.To explore the effect of changes in PDMS surface curvature on the maintenance of pluripotency of mouse embryonic stem cells,a culture matrix with different curvature was prepared to culture mouse embryonic stem cells,observe the biological morphology of cells,and detect the pluripotency of embryonic stem cells by immunofluorescence and alkaline phosphatase staining.It was found that the concave basement and the convex basement had positive effects on the pluripotency maintenance of embryonic stem cells compared with the planar basement,which could help maintain totipotency,but the concave basement had better effect.It is of great significance to the research and clinical application of embryonic stem cells.