Primary culture of fibroblast cells from rat/mouse - (Apr/11/2013 )

Dear All,
Does anyone have the protocol/paper to culture primary fibroblast cells from the tail or whatever sources of organs/tissues from rat and mouse.
Thank you.

wdyeo

I don't have one handy but the general procedure is:

Take tissue sample, sterilize surface in bleach (10s) then rinse in sterile water and rinse with 70% ethanol.
Mince tissue into small chunks using sterile scapels (may help to put softer tissues through a cell strainer).
Digest chunks with a protease for some time (tissue dependent).
Neutralize protease and plate out.

Bob, just to make sure. Do you mean to sterilize the surface of the tissue sample?

Sorry, yes, the tissue surface, this is especially important for bits with skin and hair on them, as these are typically covered in bacteria, yeasts and other fungi.

Hi bob1,
Thanks alot for the suggestion...Maybe gotta optimise own protocol