Generation and Characterization of IPSC-derived Renal Proximal Tubule-like Cells with Extended Stability

Overview

Journal Sci Rep

Specialty Science

Date 2021 Jun 3

PMID 34078926

Citations 15

Authors

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

Affiliations

Soon will be listed here.

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.

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