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