Elasticity and Topography-Controlled Collagen Hydrogels Mimicking Native Cellular Milieus bioRxiv, 2019 Jul 18

"Responses of cells grown on collagen hydrogels prepared by ET-RaM to ECM-mimicking cues. (a) Confocal fluorescence images of nuclear (blue) and actin (red) staining in MDCK cells grown on hydrogels with a flat surface or with 5-μm microgrooves and on a conventional polystyrene (PS) cell culture dish. The hydrogel and PS surfaces were visualized using fluorescent microspheres (green). (b–d) Cell area (b), morphological aspect ratio (c), and orientation (d) of MDCK cells on hydrogels with various elasticity ( E ) values and a flat surface ( T : −) or 5-μm microgrooves ( T : +) and in a PS dish are shown. Inserted bars in the graphs indicate the mean values. (e) Actin cytoskeleton beneath the nucleus (red) and (f) actin orientation. Inserted bars in the graphs indicate the mean values. Cells on 7-kPa hydrogels were excluded from analyses due to their multi-layer complexity. Statistical significance in (b–d, f) was assessed with Welch’s t test. ns, not significant (P ≥ 0.05); **P < 0.01, ***P < 0.001 (Table S2, Supporting Information). (g, h) Cardiomyocytes (g) and <t>C2C12</t> myotubes (h) grown on collagen hydrogels prepared by ET-RaM (236 kPa, 5-μm microgrooves) with nuclear (blue) and sarcomeric α-actinin (green) (g) and nuclei (blue) and actin (red) (h) staining. "