Whole Kidney primary cell culture - (Apr/20/2010 )
Does anyone have a protocol for whole kidney cell culture.
I have mouse kidneys from which I want to start primary cells.
Any input will be greatly appreciated.
This paper describes culture of explanted embryonic kidneys - is that what you mean by whole kidney cell culture?
"We cultured E11.5 ICR mouse (Taconic, Hudson, NY, USA) kidney explants for upwards of 72 h with 10 µM NRAGE morpholino (6) and 4 µM Endo-Porter. Explants were placed onto 40 µm polycarbonate filters (Millipore, Billerica, MA, USA) and cultured in a Netwell well (Corning, NY, USA). The wells were transferred to a 12-well plate with Richter’s Modified IMEM (Invitrogen, Carlsbad, CA, USA) supplemented with 10 µg/mL of Holo-transferrin (Sigma-Aldrich, St. Louis, USA) to a total volume of 750 µl. This arrangement allowed for diffusion of the culture media into the kidney explant through the filter without submerging the explant. The 12-well dish was placed in a humidified incubator at 37°C and 5% CO2 for the times described."