to get rid of differentiated cells

Hi

I am growing stem cells. But after a while i have a mixed culture of both differentiated and undifferentiated population. could you kindly tell me as to how can i get rid of the differentiated cells. i need only undifferentiated stem cells for my work

thanx for help

deb

Could you give a little more information about what cells you are growing? ES cells? Mouse or Human or..?

i am growing mouse ES cells and they are without MEFs. the number of ES colonies are getting lesser as the cells are getting differentiated though the medium has LIF. all your suggestions would be very helpful..

Is it possible to thaw a new batch of cells? Probably not the answer you want.

IS this a standard line used in many labs, or a novel/less well caracterized line?

Hi Deb,

If you need ONLY stem cells, then you're going to have to use FACS sorting, 2 rounds of it is often used in literature to make absolutely sure your sample is pure.

If you're just trying to reduce the number of differentiated cells, passing is the first step....  beyond that, perhaps try seeding to many new clonogenic colonies--and select one that has few enough undifferentiated cells.

Cheers
Luke