Figure 6.

Mechanical properties of HELP-SPAAC hydrogels influence cell spreading and morphology of hMSCs, HUVECs, and hNPCs. A,C,E) Representative images of cell morphology (hMSCs, HUVECs, and hNPCs, respectively) within HELP-SPAAC hydrogels (formulated with PEG₅AZ) with varying temperature profiles after 7 days. Nuclei staining with DAPI shown in blue, and F-actin staining with phalloidin shown in red. B,D,F) Quantification of actin area after 7 days in HELP-SPAAC hydrogels with varying temperature profiles for hMSCs, HUVECs, and hNPCs, respectively. In B) and F), actin area is normalized per cell using the DAPI counter-stain; while in D), actin area per multicellular cluster is reported (see Methods). Kruskal-Wallis with Dunn’s multiple comparisons test was used for data analysis: **p < 0.01, ****p < 0.0001. For B), n = 1,063, 823, and 899 (I, II, and III) cells examined over N = 3 independent gel samples. For D), n = 386, 448, and 503 (I, II, and III) cells examined over N = 3 independent gel samples. For F), n = 3,357, 3,013, and 3,067 (I, II, and III) cells examined over N = 3 independent gel samples.