Organoid tissue engineering

An in vitro method for obtaining an organoid having a pre-determined tissue shape and pattern, comprising:

• i. seeding stem cells capable of differentiating to form the organoid in a three-dimensional (3D) structure within a macroscopic block of hydrogel, wherein said stem cells are isolated epithelial stem cells, and the 3D structure is a cavity within the macroscopic block of hydrogel,
• wherein the cavity forms a tube comprising two open ends that allow perfusion with liquid, and,
• wherein the tube comprises a longitudinal section along the longitudinal axis of the tube, wherein the section has a rectangular cross section, wherein the rectangle has sides that are between 10 μm and 5 mm in length, or the section has a circular or elliptical cross-section, wherein the ellipse has two principal axes between 10 μm and 5 mm in length,
• ii. culturing the stem cells under self-renewal conditions, wherein the stem cells proliferate and pattern to form a colony having the same 3D structure as the macroscopic block of hydrogel and cover the 3D structure in the macroscopic block of hydrogel, and
• iii. culturing the 3D structure of step (ii) covered with the colony of the stem cells in the presence of differentiation conditions such that the colony of the stem cells undergoes morphogenesis to form the organoid with reproducible and predictable shape and pattern,
• wherein the macroscopic block of hydrogel has a minimum elastic modulus of at least 450 Pa.