Long-Term Culture of Nephron Progenitor Cells Ex Vivo
- 799 Downloads
Nephrons differentiate from the cap mesenchyme of the fetal kidney. Nephron progenitor cells that populate the cap mesenchyme efficiently balance self-renewal and epithelial differentiation to enable repeated rounds of nephron formation during development. Here we describe a method to isolate and propagate these cells from the embryonic mouse kidney. Using this method, nephron progenitor cells from a single litter of mice can be propagated to hundreds of millions of cells that express appropriate markers of the undifferentiated state and retain epithelial differentiation capacity in vitro.
Key wordsKidney Organogenesis Primary cell culture Stem cells
Research reported in this publication is supported by the National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Diseases under awards R01DK078161 (L.O.) and R24DK106743 (L.O.). Core facilities support was provided by the Maine Medical Center Research Institute core facilities for Molecular Phenotyping and Progenitor Cell Analysis [supported by the National Institute of General Medical Sciences (NIGMS) 5P30 GM106391]. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
- 1.Kobayashi A, Valerius MT, Mugford JW, Carroll TJ, Self M, Oliver G, McMahon AP (2008) Six2 defines and regulates a multipotent self-renewing nephron progenitor population throughout mammalian kidney development. Cell Stem Cell 3(2):169–181. https://doi.org/10.1016/j.stem.2008.05.020CrossRefPubMedPubMedCentralGoogle Scholar
- 2.Boyle S, Misfeldt A, Chandler KJ, Deal KK, Southard-Smith EM, Mortlock DP, Baldwin HS, de Caestecker M (2008) Fate mapping using Cited1-CreERT2 mice demonstrates that the cap mesenchyme contains self-renewing progenitor cells and gives rise exclusively to nephronic epithelia. Dev Biol 313(1):234–245. https://doi.org/10.1016/j.ydbio.2007.10.014CrossRefPubMedGoogle Scholar
- 7.Brown AC, Blank U, Adams DC, Karolak MJ, Fetting JL, Hill BL, Oxburgh L (2011) Isolation and culture of cells from the nephrogenic zone of the embryonic mouse kidney. J Vis Exp 50. https://doi.org/10.3791/2555
- 8.Ng ES, Davis R, Stanley EG, Elefanty AG (2008) A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies. Nat Protoc 3(5):768–776. https://doi.org/10.1038/nprot.2008.42CrossRefPubMedGoogle Scholar