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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Biomaterials. 2019 Feb 23;202:26–34. doi: 10.1016/j.biomaterials.2019.02.021

Figure 2. Polyacrylamide hydrogels with enhanced Matrigel conjugation induce comparable levels of hPSC attachment, proliferation, and maintenance of pluripotency regardless of substrate stiffness.

Figure 2.

a Bright-field image of H9 cells attached to Matrigel-coated hydrogel substrates (3, 14, 38 kPa) and tissue culture plastic (TCP) at day 1 in growth medium (mTeSR). All hydrogel groups enabled robust H9 cell attachment. Scale bar: 100 μm. b Effect of substrate stiffness on cell proliferation. Changes in amount of total DNA content over time confirm comparable H9 cell attachment and proliferation across three stiffnesses. Data are mean values; error bars are standard error of the mean. Groups did not significantly differ according to two-way ANOVA by Bonferroni’s multiple comparisons test. (n=3, n.s. not significant) c Immunostaining of Ki-67, a marker of actively proliferating cells, revealed that H9 cell proliferation was not influenced by substrate stiffness (day 2, mTeSR). Scale bar: 50 μm. d Expression of pluripotency markers (Oct4, Tra1-60) was maintained regardless of substrate stiffness (day 2, mTeSR). Scale bar: 100 μm. e Regardless of substrate stiffness, actin robustly polymerized into filaments and YAP translocated into the nucleus. Scale bar: 20 μm.