Figure 2.

Establishing a Tumor Organoid Model with ECM Dense Stroma

(A) Culture molds were produced by additive printing a negative mold then deposited PDMS was cured to generate a 6-well plate microwell insert. The cell-collagen type I solution was deposited, with a suspended preformed HCT-116 spheroid, into microwells and allowed to gel. Culture media, with or without 10 ng/mL of TGF-β was added to the culture media after the organoid gelated and allowed to grow for up to 7 days before being harvested. Organoids cultured in TGF-β, right, underwent sizable contraction compared to organoids in control media, center, and collagen-only controls, left.

(B) Organoid diameter was measured over 96 hr in culture conditions. Graphs represent mean ± s.e.m (n = 4).

(C) Organoid stiffness was measured over 96 hr in culture using a rheometer to determine the young's modulus of the construct. Graphs represent mean ± s.e.m (n = 4) (∗∗p ≤ 0.01; ∗∗∗∗p ≤ 0.0001).