How to make 3T3 mouse fibroblast cell bank? - (Jun/28/2006 )

Is there anybody who will guide me about basic technique to grow 3T3 fibroblasts Swiss albino obtained from Sigma to make a cell bank? I am beginner in cell culture thats why i need to know fundamentals of skin cell culture.

Are you getting skin for explants or are you getting already established lines?

For explants, cut off the epidermis. Chop the remaining tissue into small pieces and place a piece of tissue/drop of DMEM (see below). Leave for 2 days and then change the media every 2 days. Will start to see cells growing out in about a week.

For already established lines...Fibroblasts are very easy to grow.

Require DMEM/10% FBS/pen-strep in a 37oC incubator and 5% CO2.

Once they are confluent in a T 75 flask split and freeze half