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.
| Original language | English |
|---|---|
| Article number | 11575 |
| Pages (from-to) | 1-17 |
| Number of pages | 17 |
| Journal | Scientific Reports |
| Volume | 11 |
| Early online date | 2 Jun 2021 |
| DOIs | |
| Publication status | Published - Dec 2021 |
Bibliographical note
Funding Information:We would like to thank Ruthmila Paskel for her technical assistance. The research leading to these results has received support from the by the in3 project, a Marie Sklodowska-Curie Action – Innovative Training Network, under Grant No. 721975 (to PJ) and the Innovative Medicines Initiative Joint Undertaking under grant agreement no 115439, Stem-BANCC, (to PJ and GG) resources of which are composed of financial contribution from the European Union Seventh Framework Programme (FP7/2007-2013) and EFPIA. Further support was from the EU-ToxRisk project (An Integrated European “Flagship” Program Driving Mechanism-Based Toxicity Testing and Risk Assessment for the 21st Century), funded by the European Commission under the Horizon 2020 Programme (Grant Agreement No. 681002) (to PJ and JK). This publication reflects the author’s views and neither the IMI JU nor EFPIA nor the European Commission are liable for any use that may be made of the information contained therein.
Publisher Copyright:
© 2021, The Author(s).
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
Funding
We would like to thank Ruthmila Paskel for her technical assistance. The research leading to these results has received support from the by the in3 project, a Marie Sklodowska-Curie Action – Innovative Training Network, under Grant No. 721975 (to PJ) and the Innovative Medicines Initiative Joint Undertaking under grant agreement no 115439, Stem-BANCC, (to PJ and GG) resources of which are composed of financial contribution from the European Union Seventh Framework Programme (FP7/2007-2013) and EFPIA. Further support was from the EU-ToxRisk project (An Integrated European “Flagship” Program Driving Mechanism-Based Toxicity Testing and Risk Assessment for the 21st Century), funded by the European Commission under the Horizon 2020 Programme (Grant Agreement No. 681002) (to PJ and JK). This publication reflects the author’s views and neither the IMI JU nor EFPIA nor the European Commission are liable for any use that may be made of the information contained therein.
| Funders | Funder number |
|---|---|
| EU-ToxRisk | |
| Horizon 2020 Framework Programme | 681002, 721975 |
| European Federation of Pharmaceutical Industries and Associations | |
| European Commission | |
| Seventh Framework Programme | |
| Innovative Medicines Initiative | 115439 |
