Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers

Theresa Chow; Frances T M Wong; Claudio Monetti; Andras Nagy; Brian Cox; Ian M Rogers

doi:10.1038/s41536-020-0092-5

Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers

NPJ Regen Med. 2020 Apr 20:5:7. doi: 10.1038/s41536-020-0092-5. eCollection 2020.

Authors

Theresa Chow^{1

2}, Frances T M Wong², Claudio Monetti¹, Andras Nagy^{1

3

4}, Brian Cox^{2

3}, Ian M Rogers^{1

2

3}

Affiliations

¹ 1Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON Canada.
² 2Department of Physiology, University of Toronto, Toronto, ON Canada.
³ 3Department of Obstetrics and Gynaecology, University of Toronto, Toronto, ON Canada.
⁴ 4Institute of Medical Science, University of Toronto, Toronto, ON Canada.

Abstract

In order to harness the potential of pluripotent stem cells, we need to understand how to differentiate them to our target cell types. Here, we developed a protocol to differentiate mouse embryonic stem cells (ESCs) to renal progenitors in a step-wise manner. Microarrays were used to track the transcriptional changes at each stage of differentiation and we observed that genes associated with metanephros, ureteric bud, and blood vessel development were significantly upregulated as the cells differentiated towards renal progenitors. Priming the ESCs and optimizing seeding cell density and growth factor concentrations helped improve differentiation efficiency. Organoids were used to determine the developmental potential of the renal progenitor cells. Aggregated renal progenitors gave rise to organoids consisting of LTL+/E-cadherin+ proximal tubules, cytokeratin+ ureteric bud-derived tubules, and extracellular matrix proteins secreted by the cells themselves. Over-expression of key kidney developmental genes, Pax2, Six1, Eya1, and Hox11 paralogs, during differentiation did not improve differentiation efficiency. Altogether, we developed a protocol to differentiate mouse ESCs in a manner that recapitulates embryonic kidney development and showed that precise gene regulation is essential for proper differentiation to occur.

Keywords: Embryonic stem cells; Stem-cell differentiation.