Figure 3.

Generation of human kidney organoids from hPSC

(A) Timeline of kidney organoid generation from hPSCs. PPS: posterior primitive streak; IM: intermediate mesoderm; NPC: nephron progenitor cell; RV: renal vesicle; ACT: activin A; Hep: heparin.

(B) Representative bright field images showing morphological changes in the cell monolayer from day 0 to day 3 of differentiation.

(C) Representative bright field image of the cell monolayer at day 4 of differentiation showing a very compact and uniform appearance.

(D) Example of a failed formation of a day 4 cell monolayer. Yellow asterisks indicate areas of loose clusters of cells or empty areas.

(E) Representative bright field image of a day 6 spheroid (2 days after intermediate mesoderm-committed cell aggregation into spheroids). At this stage, cell spheroid is characterized by a well-compacted round morphology.

(F) Example of a failed cell aggregation in a day 6 spheroid. Note that edges are broken and disaggregated.

(G) Representative bright field image of a day 9 organoid that begin to show the formation of RVs in the organoid edges.

(H) Representative bright field image of a day 11 RV-stage organoid containing many RVs.

(I) Example of a failed formation of RVs in a day 11 kidney organoid.

(J) Representative bright field image of a day 16 kidney organoid containing multiple nephron-like structures.

(K) Examples of inefficient formation of nephron-like structures within day 16 kidney organoids. Scale bars in (B–K), 100 μm.