1 reply to this topic

[Cstar1]

Hi

I am trying to recover PBMCs from liquid Nitrogen storage for antigen stimulation work.
Do I need to rest the cells overnight before adding the antigens into the cell culture?
Because I found some of the cells don't really response to the anti-CD3 (positive control).

Thanks

[Astarte Biologics]

Cstar1, on Jul 2 2009, 03:04 AM, said:

Hi

I am trying to recover PBMCs from liquid Nitrogen storage for antigen stimulation work.
Do I need to rest the cells overnight before adding the antigens into the cell culture?
Because I found some of the cells don't really response to the anti-CD3 (positive control).

Thanks

Hi,

I've had good luck adding antigens right after thawing.  When you say that some cells don't respond to anti CD3, how are you measuring that?  Is it that some vials of cells don't respond or are you measuring by CFSE and seeing a bright population?  If you can't get any proliferation to anti CD3 there's a problem all right but it depends a bit on the details of your method.