Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability

Abstract The renal proximal tubule is responsible for re-absorption of the majority of the glomerular filtrate and its proper function is necessary for whole-body homeostasis. Aging, certain diseases and chemical-induced toxicity are factors that contribute to proximal tubule injury and chronic kidn...

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Autores principales:	Vidya Chandrasekaran, Giada Carta, Daniel da Costa Pereira, Rajinder Gupta, Cormac Murphy, Elisabeth Feifel, Georg Kern, Judith Lechner, Anna Lina Cavallo, Shailesh Gupta, Florian Caiment, Jos C. S. Kleinjans, Gerhard Gstraunthaler, Paul Jennings, Anja Wilmes
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cabcf3e0db84468c83c49d606b6f2e22
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spelling	oai:doaj.org-article:cabcf3e0db84468c83c49d606b6f2e222021-12-02T18:24:54ZGeneration and characterization of iPSC-derived renal proximal tubule-like cells with extended stability10.1038/s41598-021-89550-42045-2322https://doaj.org/article/cabcf3e0db84468c83c49d606b6f2e222021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89550-4https://doaj.org/toc/2045-2322Abstract The renal proximal tubule is responsible for re-absorption of the majority of the glomerular filtrate and its proper function is necessary for whole-body homeostasis. Aging, certain diseases and chemical-induced toxicity are factors that contribute to proximal tubule injury and chronic kidney disease progression. To better understand these processes, it would be advantageous to generate renal tissues from human induced pluripotent stem cells (iPSC). Here, we report the differentiation and characterization of iPSC lines into proximal tubular-like cells (PTL). The protocol is a step wise exposure of small molecules and growth factors, including the GSK3 inhibitor (CHIR99021), the retinoic acid receptor activator (TTNPB), FGF9 and EGF, to drive iPSC to PTL via cell stages representing characteristics of early stages of renal development. Genome-wide RNA sequencing showed that PTL clustered within a kidney phenotype. PTL expressed proximal tubular-specific markers, including megalin (LRP2), showed a polarized phenotype, and were responsive to parathyroid hormone. PTL could take up albumin and exhibited ABCB1 transport activity. The phenotype was stable for up to 7 days and was maintained after passaging. This protocol will form the basis of an optimized strategy for molecular investigations using iPSC derived PTL.Vidya ChandrasekaranGiada CartaDaniel da Costa PereiraRajinder GuptaCormac MurphyElisabeth FeifelGeorg KernJudith LechnerAnna Lina CavalloShailesh GuptaFlorian CaimentJos C. S. KleinjansGerhard GstraunthalerPaul JenningsAnja WilmesNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Vidya Chandrasekaran Giada Carta Daniel da Costa Pereira Rajinder Gupta Cormac Murphy Elisabeth Feifel Georg Kern Judith Lechner Anna Lina Cavallo Shailesh Gupta Florian Caiment Jos C. S. Kleinjans Gerhard Gstraunthaler Paul Jennings Anja Wilmes Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability
description	Abstract The renal proximal tubule is responsible for re-absorption of the majority of the glomerular filtrate and its proper function is necessary for whole-body homeostasis. Aging, certain diseases and chemical-induced toxicity are factors that contribute to proximal tubule injury and chronic kidney disease progression. To better understand these processes, it would be advantageous to generate renal tissues from human induced pluripotent stem cells (iPSC). Here, we report the differentiation and characterization of iPSC lines into proximal tubular-like cells (PTL). The protocol is a step wise exposure of small molecules and growth factors, including the GSK3 inhibitor (CHIR99021), the retinoic acid receptor activator (TTNPB), FGF9 and EGF, to drive iPSC to PTL via cell stages representing characteristics of early stages of renal development. Genome-wide RNA sequencing showed that PTL clustered within a kidney phenotype. PTL expressed proximal tubular-specific markers, including megalin (LRP2), showed a polarized phenotype, and were responsive to parathyroid hormone. PTL could take up albumin and exhibited ABCB1 transport activity. The phenotype was stable for up to 7 days and was maintained after passaging. This protocol will form the basis of an optimized strategy for molecular investigations using iPSC derived PTL.
format	article
author	Vidya Chandrasekaran Giada Carta Daniel da Costa Pereira Rajinder Gupta Cormac Murphy Elisabeth Feifel Georg Kern Judith Lechner Anna Lina Cavallo Shailesh Gupta Florian Caiment Jos C. S. Kleinjans Gerhard Gstraunthaler Paul Jennings Anja Wilmes
author_facet	Vidya Chandrasekaran Giada Carta Daniel da Costa Pereira Rajinder Gupta Cormac Murphy Elisabeth Feifel Georg Kern Judith Lechner Anna Lina Cavallo Shailesh Gupta Florian Caiment Jos C. S. Kleinjans Gerhard Gstraunthaler Paul Jennings Anja Wilmes
author_sort	Vidya Chandrasekaran
title	Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability
title_short	Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability
title_full	Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability
title_fullStr	Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability
title_full_unstemmed	Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability
title_sort	generation and characterization of ipsc-derived renal proximal tubule-like cells with extended stability
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/cabcf3e0db84468c83c49d606b6f2e22
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Generation and characterization of iPSC-derived renal proximal tubule-like cells with extended stability

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