Figure 2.

Differentiation of hPSCs into Kidney Organoids

(A) Schematic of the protocol.

(B) Bright-field images of ~50% confluent hPSC colonies on day 0.

(C) High magnification of desired appearance of hPSC colonies after Dispase-treatment (6 min at 37˚C) showing the edge of the colony starting to curl up (white arrowhead).

(D) Upon overdigestion with Dispase (shown here is 12 min at 37˚C) the colonies are peeling off the growth surface (red arrowheads) and detach (red arrows), leading to increased cell death and low efficiency in organoid formation.

(E) Fragments of hPSC colonies in suspension after Dispase digest.

(F and F′) Day 2 embryoid bodies.

(G) Tilted 6-well plate for day 2 half-medium change.

(H) Orbital shaker used from day 2 onward.

(I and I′) Day 3 embryoid bodies.

(J) A spinning bioreactor can be used for embryoid body / organoid culture from day 3 onward.

(K) Tubule formation is visible on the surface of day 8 organoids (arrows in K′).

(L) Day 12 kidney organoids.

Scale bars, 1 mm (B, F, and I); 400 μm (C−E, F′, I′, K, and L).