T47D culture

[Anuska]

Do somebody have an explanation about the different behabior when culturing cells in flasks. I have lots of cell lines that canstand a lot of passages in the same flask, but the T47D and modified T47D that can't be trypsinated in the same flask for longer than 3 passages. Cells start to form clusters, or detatach from the flask.

thanks

[satyamka]

hello dear
I would like to suggest you this:- try to trypsinise in sub-confluent stage and after trpsinising cells, just make sure that after harvesting a fraction of cells/seeding in other flasks, you have made a single cell suspension. As in case the cells after trypsinisation will be in clusters/groups, they will make small colonies, and at that very plave, due to excessive growth, there may be cell detachment, as they will not get space for monolayer formation.
thanks
Saty

[itsmadhvi]

I agree with Satyamka, I m handling T47D routinely, this cells have a tendency to grow in clumps, so it is very neccessary for you to homogenise them properly after trypsinization. Also before trypsinization include one or two PBS washes.