Figure 4.

Substrate Regulates Cell Shape and Gene Expression

(A) Morphology change of MCoG cells and DCoG cells on different substrates during long-term passage. In each condition, the left panel is the phase contrast image and the right panel is the SEM image.

(B) Immunofluorescence images of single AIT and AST cells on the MG and GNF substrates, respectively. White arrows indicate cells engaged in spreading.

(C) Fraction of detached cells plotted as a function of hydrodynamic pressure P. Data points were fitted with the cumulative distribution function of normal distribution, and the critical pressure P^∗ was determined as the required pressure at which 50% of cells were detached (mean ± SE, n ≥ 500 cells). Four conditions are investigated: AIT cells on MG (orange), AST cells on MG (red), AIT cells on GNF (green), and AST cells on GNF (blue).

(D) Relative expression of hPSC-specific genes in AIT and AST cells on MG and GNF substrates (mean ± SD, n ≥ 3 independent experiments, ^∗p < 0.05, ^∗∗p < 0.01). To avoid the interference of ROCK inhibitor on cell adhesion, data in this figure were obtained more than 48 hr after withdrawal of the ROCK inhibitor from cell cultures.

See also Figure S5.