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Whole Kidney primary cell culture


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#1 Sank

Sank

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Posted 20 April 2010 - 07:09 AM

Hello everyone
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.

Regards
Sankella

#2 Jon Moulton

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Posted 20 April 2010 - 12:42 PM

This paper describes culture of explanted embryonic kidneys - is that what you mean by whole kidney cell culture?

http://www.ncbi.nlm....les/PMC2646631/

"We cultured E11.5 ICR mouse (Taconic, Hudson, NY, USA) kidney explants for upwards of 72 h with 10 M NRAGE morpholino (6) and 4 M Endo-Porter. Explants were placed onto 40 m 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 Richters Modified IMEM (Invitrogen, Carlsbad, CA, USA) supplemented with 10 g/mL of Holo-transferrin (Sigma-Aldrich, St. Louis, USA) to a total volume of 750 l. 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 37C and 5% CO2 for the times described."
Jon D. Moulton
Gene Tools, LLC
www.gene-tools.com




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