Freezing down cells

Add up to 0.9mLs of cells in media to cryovial. Add equal volume of freezing media, cap and invert 2-3x to mix. Place in Mr. Frosty and store at -80C for at least 4 hours before transferring to a box at -80 or to liquid nitrogen storage.

• Bone marrow: No more than 107 cells per mL.

Freezing media

• 20% DMSO
• 80% serum (BGS, FBS)

Thawing cells

Thaw vials of cells in warm tap water until liquid. Gently remove cells to a tube with pre-warmed media, rinse the cryovial once with media. Pellet cells gently in centrifuge (1000 rpm for 5min) and remove supernatant. Resuspend gently with media and plate, etc.