Modulating the Substrate Stiffness to Manipulate Differentiation of Resident Liver Stem Cells and to Improve the Differentiation State of Hepatocytes
Soft substrate induces a rapid and homogeneous epithelial differentiation of RLSCs. (a) Phase-contrast micrographs of RLSCs grown on Petri plastic dish (CTRL; GPa) and on hydrogels with kPa and 80 kPa, for 24 and 48 hours. Images are representative of three independent experiments. Scale bar: 100 μm. (b) Phase-contrast micrographs and immunofluorescence of cells cultured on plastic (CTRL), 0.4 kPa and 80 kPa for 24 and 48 hours, stained for HNF4α, Vimentin, and E-cadherin. The nuclei were stained with DAPI. Images are representative of three independent experiments. Scale bar: 50 μm. (c) Western blot analysis of Vimentin at 24 and 48 hours after seeding on substrates with the indicated values. CDK4 was used as a loading control.