I'm working with BJAB lymphocytic cell line, culturing them with RPMI media (with L-glutamine) + 10%FBS + 1% antibiotics. After two days of changing media by centrifugation, they started to adhere to the T flask and a monolayer has formed. Does someone know what could be happening? Is the media lacking something or should it be a step in the handling that I'm missing?
Sandra Cortes you should check the type of culture flask first that should not be tissue-culture treated one, and maintain cell culture with requires agitation (i.e., shaking or stirring) for adequate gas exchange. Try this maybe this can help you.
Do you keep the flask in a horizontal or vertical position in the incubator? If you are using a tissue culture treated flask, I recommend that you keep your flask in a vertical position in the incubator.
I second Sameer Varma 's comment regarding the flask type. It's not the end of the world if you are indeed using a tissue-culture treated flask for suspension cells. The cells will usually detach if you wash the surface with media several times when you are splitting.
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