I need a protocol for harvesting fibroblasts from Mouse tail tips - (Mar/12/2010 )
I need a protocol for harvesting fibroblasts from Mouse tail tips. The mice are post natal p15-30.
Can anyone of you provide me with a protocolto harvest them, especially in what kind of media do they growth and what kind of surface coating they need?
I usually take the skin away from the tail and then cut it into pieces inside one well of a 6 well plate (not coated with anything). I leave the pieces for a few minutes till they stick to the plate. Then I add 2 ml advanced DMEM (5% FCS, Pencillin/ Strptomycin, Glutamin) to the wells without dislodging the pieces and put the plate in the incubator. A layer of fibroblasts should grow out in a few days. Once they are dense enough, I split them and transfer them to bigger plates.