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 media used for in vitro growth of mouse nephron progenitors and 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 preservation of uninduced and induced cap mesenchyme as well as a transiting mesenchymal-epithelial state. Concomitantly, differentiating and differentiated epithelial lineages are also maintained.
These data, showing in-vitro expansion of discrete stages of human nephrogenesis at single-cell resolution may be used for drug screening on a full repertoire of developing kidney cells and possibly prospective isolation of mesenchymal or epithelial renal lineages for regenerative medicine.
