Figure 1.

HELP matrix for patient‐derived intestinal organoid formation, growth, and passaging. a) Enteroids were generated from intestinal tissue biopsies from human patients. b) Schematic of HELP matrix, which is composed of benzaldehyde‐modified hyaluronan (HA) and hydrazine‐modified elastin‐like protein (ELP). Hyaluronan can engage the CD44 receptors on cells, while recombinant ELP contains an RGD peptide ligand that engages cell integrin receptors. c) Schematic of enteroid passaging techniques. Enteroids can either be dissociated into single cells or directly re‐embedded as fully formed enteroids into a new material at the time of passaging. d) Brightfield and confocal fluorescence micrographs of enteroids in different materials, when dissociated (left) and re‐embedded (right) in these materials. e) Representative brightfield images of enteroids grown on Passage 1 and Passage 5 in HELP, with dissociation into single cells at the time of each passage. f) Growth curves of dissociated enteroids grown from Passage 12 in EHS matrix or Passage 12 in HELP; at each timepoint, a two‐tailed Student's t‐test was conducted, n = 3, n.s. = not significant. g) Enteroid formation efficiency for enteroids grown from single cells in EHS matrix, HELP, or ELP only. One‐way ANOVA with Tukey's multiple comparisons testing; ** = p < 0.01, n = 3, n.d. = none detected. All data shown are mean ± SD.