We would like to freeze primary myeloma cells before testing the compounds. Does anyone have any experience with this? What cryomedia did you use? Any advice on thawing the cells?
Hi Figueroa,
We freeze our primary myeloma cells in cryomedia containing 50% FBS, 40% RPMI and 10% DMSO. We thaw cells in a 37oC water bath and add cells to an up right t-25 flask containing ~20ml RPMI 1640 medium containing 20%FBS (R20). We allow the cells to settle to the bottom of the flask for 3-4 hours. We then remove ~15 ml form the flask and replace with 5-10 ml of new R20. We use this method to reduce the DMSO concentration. This significantly reduces cells death compared to centrifugation. We normally get 85-90% viability.
try 90% serum and 10% DMSO, believe me, it always works perfect. When you thaw it, try to thaw it quickly in 37 degree water bath. But when you freeze it down, try to freeze it slowly, for example, first put it in 4 degree, then -20, then -80, then liquid nitrogen. Or if you like just put it in strero box and leave it in -80, a couple of days later, relocate it in to liquid nitrogen. Good luck!
Thank you for your reply. Yesterday I tried the suggestion from Daniel Medina, so I will find out how it goes. About 90% serum and 10% DMSO, I could also have a look to it when the next sample arrives.
Primary myeloma cells don't survive that well so we normally use them straight away to test inhibitors and don't bother freezing them.
- Badges
- Science topic
- Similar topics
- Medicine
- Oncology
- Cancer Research
- Cancer Biology