Figure 4.

a) When encapsulated in hydrogels that were subsequently softened, colony viability was maintained and unaffected by irradiation for less than 10 seconds of exposure. b) To study crypt formation as a function of hydrogel moduli, ISC colonies were released from Matrigel and encapsulated in photodegradable hydrogels. After 24 hours, hydrogels were swollen with LAP and Glutathione in Flurobrite media for 30 minutes and then irradiated with 365 nm light. After irradiation, colonies were switched to differentiation media. 48 hours after softening, colonies were fixed and immunostained. c) Cell laden hydrogels were softened for 0, 2, 5, 8 or 10 seconds and crypt forming efficiency was quantified as the percentage of living colonies that formed crypts. d) Resulting crypts were measured by quantifying the distance from the main colony body to the tip of the crypt. e) Crypt length was quantified for all softening conditions and showed moduli dependence, f) as did the number of crypts that formed per colony. g) Crypts were marked by the presence of lysozyme producing Paneth cells. Scale bar 100 μm. Data is represented as means +/− standard deviation. At least three hydrogels were analyzed per condition and data was compared using a one-way Anova with multiple comparisons. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001