Rescue RAW cell culture - RAW cells past confluent point... can they be rescued? (Oct/11/2010 )

Hello,

I perform RAW cell tissue culture in a dish, using DMEM. I believe they reach confluent point every 48 hours. So I split them every other day.

Problem is... I split them last Thursday... then did not do so until today (Monday). So it's been four days. The cells look unhealthy... lots of dendrites spurting... the medium is turning faintly yellow from its original brilliant pink.

My question is: Can these cells be rescued? If I diligently split them from now on, can they revert back to their healthy, rounded shape? Or is the dendritic activated state here to stay? (Fyi: healthy RAW cells are round and same-sized. UNhealthy RAW cells change shapes and develop a ton of dendrites)

Thanks a lot for any feedback!

cookiedoughgirl on Tue Oct 12 00:36:18 2010 said:

Sorry I didn't see this yesterday. The color change is probably due to a change in pH of the media. See what happens after you give them fresh media.

I don't know much about RAW cells but my advice would be to get another vial up from ln2 if you can.

Even if you manage to "rescue" your cells by treating them well from now on, and even if they do return to their usual morphology, you won't necessarily be able to tell what other damage has been done and if anything else about them has been permenantly changed.

Hi, I didnt know that cells could not be kept above -70. I kept it at -50 to -30 for 2 weeks (because the freezer in my lab cannot decrease the temperature anymore. It just maintain from -30 to -50). After I realized it, I transferred it to liquid nitrogen. Can my cells still be revived? What are the chances? And how can I increase the viability?

By the way, the cell is RAW cells. And I havent thawed it since receiving it from ATCC.

P/S: I am new to cell culture.