Directed differentiation of human pluripotent stem sells for the generation of high-order kidney organoids

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...

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Detalles Bibliográficos
Autores: Selfa, Idoia Lucía, Gallo, Maria, Montserrat, Núria, Garreta, Elena
Tipo de recurso: capítulo de libro
Estado:Versión aceptada para publicación
Fecha de publicación:2020
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/173237
Acceso en línea:https://hdl.handle.net/2445/173237
Access Level:acceso abierto
Palabra clave:Ronyó
Cèl·lules mare
Diferenciació cel·lular
Kidney
Stem cells
Cell diferentiation
Descripción
Sumario: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.