STAR Protocols (Jun 2024)

Protocol for isolating murine kidney tubules and ex vivo cell death assays

  • Alexia Belavgeni,
  • Francesca Maremonti,
  • Andreas Linkermann

Journal volume & issue
Vol. 5, no. 2
p. 103005

Abstract

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Summary: Isolating kidney tubules offers insights into their biological function without stroma, vascular cells, and immune system interference. Our murine tubule isolation protocol focuses on ex vivo cell death assays. We describe steps for solution preparation; kidney extraction, decapsulation, and slicing; and tubule isolation. We also outline assays like western blotting, lactate dehydrogenase release assay, and live-cell imaging of vital dyes during experimental acute tubular necrosis. This adaptable protocol allows the generation of outgrown primary tubular cells that maintain the features of tubular cells. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords