Cells not growing - (Apr/27/2006 )

We recently moved to a new lab. First my cells (rat hepatoma cell line) got contaminated and then i cultured a new batch of cells which didnt get contaminated but stopped growing. Contrary to this, cells of other labmates who moved at the same time are growing well.

What should i do so that the cells grow like earlier.

Rohit

Why didnt anybody reply to my post..........damn that is so urgent and making my life miserable.

Depends on the cell line ... where you got it from? Was it from frozen or from a fresh specimen? What sort of media are you using?? Are your labmates cells in the same incubator? Perhaps more details and someone could help.

first of all, you only waited about 3 hours?

also, without more information how can we help you? you lost a batch due to contamination...unfortunately, it happens, and is more likely when the cells are moved around.

second, you lost another batch because they 'stopped growing'
this means, probably either the cells are bad, or one of the reagents is bad, or the CO2 incubator is bad, you added cold medium...there are many possible reasons for this

I am very sorry that you are so upset, but with what you've given us, how can you expect us to help?

to solve your problem, you need to slow down...take a deep breath...and try to figure out why the second batch stopped growing. (we already know why the first batch got contaminated; at some point microorganisms crawled into the dish and starting sharing space with your cells)

can you tell us any more about the second batch? color change? have you checked your media for expiration? have you checked your incubator for proper sensing of temperature of CO2 level?

get checked for mcoplasma contamination in the culture

i agree with them,there are many affect factors, why not try them one by one carefully? good luck!