Staff member


Maria Gallo

PhD Student
Pluripotency for organ regeneration
mgallo@ibecbarcelona.eu
+34 934 020 566
Staff member publications

Selfa, Idoia Lucía, Gallo, Maria, Montserrat, Nuria, Garreta, Elena, (2021). Directed differentiation of human pluripotent stem sells for the generation of high-order kidney organoids Programmed Morphogenesis: Methods and Protocols (ed. Ebrahimkhani M.R., Hislop J.), Springer US (New York, NY) 2258, 171-192

Our understanding in the inherent properties of human pluripotent stem cells (hPSCs) have made possible the development of differentiation procedures to generate three-dimensional tissue-like cultures, so-called organoids. Here we detail a stepwise methodology to generate kidney organoids from hPSCs. This is achieved through direct differentiation of hPSCs in two-dimensional monolayer culture toward the posterior primitive streak fate, followed by induction of intermediate mesoderm-committed cells, which are further aggregated and cultured in three-dimensions to generate kidney organoids containing segmented nephron-like structures in a process that lasts 20 days. We also provide a concise description on how to assess renal commitment during the time course of kidney organoid generation. This includes the use of flow cytometry and immunocytochemistry analyses for the detection of specific renal differentiation markers.