Methods and culture substrates for controlled induction of biomimetic neural tissues comprising singular rosette structures

A method of producing a plurality of biomimetic neural tissues, comprising:

• (a) seeding human pluripotent stem cells (hPSCs) in the presence of a Rho kinase inhibitor onto a micropatterned substrate, wherein the substrate is comprised of an array of micropatterned regions configured to instruct biomimetic neural morphogenesis of cells cultured thereon;
• (b) culturing the seeded cells of (a) on the micropatterned substrate for a first culture period of about one to two days in the presence of a first neural differentiation base medium to obtain a first cell population, wherein the first neural differentiation base medium comprises a Rho kinase inhibitor;
• (c) culturing the cells of (b) for a second culture period of about 2 to about 6 days under adherent culture conditions in a second neural differentiation base medium;
• and
• (d) producing a plurality of biomimetic neural tissues arrayed on the micropatterned substrate, wherein at least 75% of the arrayed tissues consist of a single polarized rosette structure in which at least 80% of cells of the structure are Pax6+/N-cadherin+ neuroepithelial cells.