PBMC-B cell - (Apr/23/2002 )

Dear Sir/Madame

I am currently undertaking a PhD work in the immunology barnch of the GIMAP laboratory in the St Etienne school of Medicine (France). My work is under the direction of Pr. C GENIN and Dr. O GARRAUD.
I am looking for a precise expérimental protocol for human B cell and PBMC culture (conditions, type of plate, cell density, reagents…) allowing me to decrease the baseline production of human Ig during cell culture – This Ig product is measured by ELISA.

I would like also to optimize culture conditions to increase Ig production.
I would be very grateful if you gave me your precious help.

Sincerely yours.

Hi there!

I'm not quite sure, but maybe it'll help. I'm doing PBMC Prep by the Greiner LeucoSep ? system (available at www.greiner-lab.com).

The isolated PBMC's are resuspended and cultured in RPMI 1640 special (containing stab. Glutamine, NEAA, 25mM HEPES, 1mM Pyruvate, 2,0 g/L NaHCO3 sterile filtered). We get this from PAN and complemenmt it with 5% of human serum.

The cells are cultured in 24 well multiwell plates at at density of 2 millon/well. Plates are from Falcon, #35 3047.

Mike

Never give up, never surrender!