Isolation and Culture of Cells from the Nephrogenic Zone of the Embryonic Mouse Kidney

Aaron C. Brown; Ulrika Blank; Derek C. Adams; Michele J. Karolak; Jennifer L. Fetting; Beth L. Hill; Leif Oxburgh

doi:10.3791/2555

Isolation and Culture of Cells from the Nephrogenic Zone of the Embryonic Mouse Kidney

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Isolation and Culture of Cells from the Nephrogenic Zone of the Embryonic Mouse Kidney

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08:06 min

April 22, 2011

DOI: 10.3791/2555-v

Aaron C. Brown¹, Ulrika Blank², Derek C. Adams¹, Michele J. Karolak¹, Jennifer L. Fetting¹, Beth L. Hill¹, Leif Oxburgh¹

¹Department of Molecular Medicine,Maine Medical Center Research Institute, ²Molecular Medicine and Gene Therapy,Lund University Hospital

In this report we describe a method for the isolation and culture of the progenitor cell niche from the embryonic mouse kidney that can be used to study signaling pathways regulating stem/progenitor cells of the developing kidney. These cultured cells are highly accessible to small molecule and recombinant protein treatment, and importantly also to viral transduction, which allows efficient manipulation of candidate pathways.

The overall goal of this procedure is to generate a primary culture of nephrogenic zone cells from the developing kidney. This is accomplished by first dissecting out kidneys from day 17.5 embryos. The next step of the procedure is to remove the kidney capsules.

A limited enzymatic digestion of the organs is then performed to dislodge cap mesenchyme and corticals stroma cells from the nephrogenic zone. The final step of the procedure is to plate the cells in fibronectin and culture in serum free medium. Ultimately, results can be obtained that demonstrate a purified nephrogenic zone cell population by immuno staining of specific markers.

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