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Kidney Week

Abstract: TH-PO0566

Novel Insights on the Origin of Kidney Stem/Progenitor Cells Derived from Neonatal Urine

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine

Authors

  • Deffrennes, Sara, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium
  • Lantermans, Hildo C., Amsterdam UMC Locatie AMC, Amsterdam, NH, Netherlands
  • Rayyan, Maissa, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium
  • Van den Heuvel, Lambertus P.W.J., Radboud Universiteit, Nijmegen, GE, Netherlands
  • Arcolino, Fanny Oliveira, Amsterdam UMC Locatie AMC, Amsterdam, NH, Netherlands
  • Levtchenko, Elena, Amsterdam UMC Locatie AMC, Amsterdam, NH, Netherlands
Background

In the developing kidney, nephrons develop from SIX2+ nephron progenitor cells (NPC). NPC are exclusively present during nephrogenesis, which is completed around 36w of gestational age (GA). We have previously described the isolation of SIX2+ kidney stem/progenitor cells from the urine of preterm neonates, named neonatal kidney/stem progenitor cells (nKSPC). Here, we aimed to determine the efficiency of nKSPC isolation across all GA and characterize them in-depth.

Methods

35 urine samples were obtained from 34 neonates across all GA categories (extreme preterm(<28w), very preterm(28–32w), moderate-late preterm(32-37w), term(>37w)). Per sample, up to 10 cell colonies were isolated to obtain clones. Residual cells from each sample were pooled together (termed ‘pool’) and cultured as well. SIX2 expression was determined and differentiation potential into kidney epithelial cells was evaluated. Thus far, 2 pools and 3 clones were analyzed using scRNAseq.

Results

28 samples yielded cell growth, yielding 124 clones and 22 pools of which 38 clones and 9 pools were SIX2+. Interestingly, SIX2+ cells were derived from all GA categories, including term. There was no difference in proliferative and differentiation potential according to GA. With scRNAseq we observed that SIX2+ clusters do not only express NPC markers but also some podocyte markers. This co-expression of SIX2 and podocyte genes was more pronounced in cells from term neonates compared to preterm, with expression of mature podocyte genes only in term samples. SIX2 negative clusters were mainly characterized by expression of markers of the developing tubule.

Conclusion

This study demonstrates that SIX2+ nKSPC can be isolated from the urine of neonates, independently of the GA at birth. The co-expression of NPC and podocyte genes suggests that nKSPC were podocytes at the time of urine collection and de-differentiated to a SIX2+ stem cell-like state during culturing. In line with our results, others have demonstrated that in the developing kidney podocyte precursors exhibit close similarity to NPC regarding their epigenetic landscape. Our study provides novel insights into human kidney development with evidence for podocyte-to-NPC plasticity, whilst also consolidating the nKSPC as a source for disease modelling and kidney regenerative medicine.

Digital Object Identifier (DOI)