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Protocol for mouse splenocytes - storage, media etc (Apr/13/2005 )

Hi

I am new in cellular immunology and therefore I really need help.

Do anyone have a working protocol for the isolation of leucocytes from mouse spleen.

I would like to get details of the media composition, hints and other things "that you just need to know".

Can the splenocytes be stored for 2 days before use, and if so, do they need som special media or do you just change the medium daily.

I expect to use the cells for ELISPOT.

Thank you very much in advance

The novice

-bobhund-

we use RPMI 1640 supplemented with 5% FCS and 10mM HEPES for our mouse splenocytes, medium is changed every day, but don't expect them to be very longlived... 1 Week is possible, though....

splenocytes are isolated by grinding the spleen on a cell strainer (100┬Ám) and lysing the erys... if you want lymphos only, you'll have to do depletion of the other cell types or positive selection of lymphos by magentic beads, for example...

mike

-jadefalcon-

Thank you very much for the fast reply.

Cheers

J

-bobhund-