How should fibroblast cell cultures be recovered from cryogenic storage?
• Prepare appropriate recovery medium (see Shipping Sheets for individual cell line). • Remove one ampule or cryovial from frozen storage and place immediately in a 37C water bath and agitate vigorously. • Once completely thawed, wipe ampule or cryovial with a 70% alcohol sponge. Score the neck of a glass ampule and open utilizing an ampule opener. • Remove the contents of the ampule or cryovial using a sterile transfer pipette and place in a T25 tissue culture flask containing 5 ml of the appropriate fresh growth medium for fibroblast cultures. • If a cell count is required, mix the contents of the flask gently with a 1 ml pipette and remove 0.2 ml for a 1:5 diluted cell count. Place the flask in the 37C incubator lying cell surface down. Gently swirl the flask to distribute the cell suspension evenly over the flask surface. Adjust the cap to allow appropriate gas exchange (depending on buffering system of the medium). Fibroblast cultures should be refed with fresh medium the day afte
