Hello!

I have been culturing GM11994- B lymphocyte cells from Coriell Institute. They are suspension cells and like to grow in colonies. However, after genetic modifications, I sorted single cells into each well in a 96-well plate. Poor viability was observed in the beginning. Then I started culturing it on feeder layers. I could see that cells started growing in colonies and had to be re-fed with feeder cells after the previous batch deteriorated. It's been around 10 weeks now but they are still in a 96-well plate and seem to have about 40% confluency only. Also, they are cultured in TC-treated plates.

I want to know,

1. does the doubling time reduce in plates compared to flasks?

2. I observed the cells are attached and is not that easy to dissociate them while transferring them to another plate. Is it because of the plates or because they are attached to feeder cells?

3. Does attachment have any change in the morphology of the cells?

It would be very helpful if I could know more about single colony formations from single cells.

Thanks in advance!