when a small frozen cell pellet remained I added dropwise pre-warmed media to the collection tube. I used the same viability/surface markers to stain for flow and the majority of cells (75%) appear to be dead. Can anyone suggest a freezing media or an alteration to my protocol in order to avoid the cell death after freeze/thawing? I used cryostor to avoid any animal serum to stop any immunological reactions. thaw them by putting them in 37C waterbath and when a small frozen cell pellet remained I added dropwise pre-warmed media to the collection tube. I used the same viability/surface markers to stain for flow and the majority of cells (75%) appear to be dead. Can anyone suggest a freezing media or an alteration to my protocol in order to avoid the cell death after freeze/thawing?

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