Murine Renal Cell Isolation: A Technique to Isolate and Generate Primary Culture of Renal Tubular Epithelial Cells

Abstract

Source: Ding, W. et al. Isolation, Characterization and High Throughput Extracellular Flux Analysis of Mouse Primary Renal Tubular Epithelial Cells. J. Vis. Exp. (2018)

In this video, we demonstrate the steps to isolate and purify renal tubular epithelial cells for a wide range of molecular biology and cell culture studies.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Tissue Processing and Primary Tubular Cells Isolation Remove the renal capsules and medulla, mince both kidneys into tiny pieces, and incubate them in 10 mL of a digestion buffer in a 37 °C oven with gentle rotation for 5 min. Remove any undigested kidney tissues by passing the buffer through a 70 μm…

Materials

Collagen I Rat Protein, Tail	ThermoFisher Scientific	A1048301
Collagenase Type II	Worthington	LS004176
Renal Epithelial Cell Growth Medium 2 Kit	PromoCell	C-26130
TC10 automated cell counter	Bio-Rad	506BR2119