Isolation of T4 cells from xenograft tumors
Put mouse down
Wash skin by dipping mice into ETOH
Remove tumor and put into 5ml PBS containing Fungizone and Penstrep
Swirl around to rinse tumor and pour on to a sterile plate
Remove tumor from PBS place on sterile plate or lid
Remove any skin-rinse a second time if needed
Chop into fine pieces with scalpel blades
Transfer using blade into 2ml H14 media containing Fungizone and Penstrep
Spin down 800rpm
Remove media leaving ~300ul
Add 2ml .25% trypsin and place on rocker for 5min
Spin 800rpm for 5min
Resuspend in DMEM/F12 with10% FCS
Spin 800rpm for 5min
Resuspend in H14 media containing Fungizone and Penstrep
Plate onto vitrogen coated flask of plate
(volume of media and size of flask depend on tumor size I used a 625px2 flask)
change media the next day and keep close eye out for contamination