20311.3 Alternative Protocols for Directing Differentiation
of hPSCs to the Kidney
Alongside the identification of this protocol for the generation of complex kidney
organoids, a number of other protocols for the renal differentiation of hPSC in vitro
have been reported. In most cases, these directed differentiation protocols also
involve stepwise differentiation through embryological stages leading to kidney
development. Hence, this again requires the derivation of the posterior PS, followed
by IM and then MM and/or UB. Other published protocols demonstrate the
Fig. 11.3 Cells present within kidney organoids. Kidney organoids generated via directed dif-
ferentiation of human pluripotent stem cells contain multiple cellular components reflecting the
cell types present during kidney morphogenesis. Organoids (a) contain collecting duct epithelium
(b) positive for Dolichos biflorus agglutinin (DBA) together with patterning and segmenting neph-
rons (c), including distal tubules expressing CDH1 (d) and proximal tubules expressing MEGALIN
and CUBILIN (CUBN), and positive for Lotus tetragonolobus lectin (LTL) (e, f) and podocytes
within maturing glomeruli (g–i). Intact glomeruli can be sieved from organoids (g) and contain
tightly packed podocytes (h) positive for SYNAPTOPODIN (SYNPO) and CD2AP (i). Surrounding
the nephrons, early endothelial cells (SOX17 + CD31+) and perivascular cells (PDGFRA+) form (j,
k), with some beginning to enter the forming glomeruli (l) (Adapted from Takasato et al. 2015 with
additional images from Jessica Vanslambrouck and Lorna Hale)
11 Recapitulating Development to Generate Kidney Organoid Cultures