Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level

Naomi Pode-Shakked, Rotem Gershon, Gal Tam, Dorit Omer, Yehudit Gnatek, Itamar Kanter, Sarit Oriel, Guy Katz, Orit Harari-Steinberg, Tomer Kalisky, Benjamin Dekel

Research output: Contribution to journalArticlepeer-review

Abstract

During nephrogenesis, stem/progenitor cells differentiate and give rise to early nephron structures that segment to proximal and distal nephron cell types. Previously, we prospectively isolated progenitors from human fetal kidney (hFK) utilizing a combination of surface markers. However, upon culture nephron progenitors differentiated and could not be robustly maintained in vitro. Here, by culturing hFK in a modified medium used for in vitro growth of mouse nephron progenitors, and by dissection of NCAM+/CD133 progenitor cells according to EpCAM expression (NCAM+/CD133/EpCAM, NCAM+/CD133/EpCAMdim, NCAM+/CD133/EpCAMbright), we show at single-cell resolution a preservation of uninduced and induced cap mesenchyme as well as a transitioning mesenchymal-epithelial state. Concomitantly, differentiating and differentiated epithelial lineages are also maintained. In vitro expansion of discrete stages of early human nephrogenesis in nephron stem cell cultures may be used for drug screening on a full repertoire of developing kidney cells and for prospective isolation of mesenchymal or epithelial renal lineages for regenerative medicine.

Original languageEnglish
Pages (from-to)279-291
Number of pages13
JournalStem Cell Reports
Volume9
Issue number1
DOIs
StatePublished - 11 Jul 2017

Keywords

  • Wilms’ tumor
  • cancer stem cells
  • kidney stem/progenitor cells
  • renal development
  • single cell gene expression analysis
  • stem cell markers
  • stem cells

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