I would like to expand memory B cells from a single memory B cell, and we are working on establishing hCD40L expressing -3T3 or -293T cell lines. Before we get the hCD40L-cell lines, we have tried culturing memory B cells without feeder cells in RPMI 1640 supplemented with 20% HI FBS, 50 ng/ml IL21, 10 ng/ml IL4, 20ng/ml CD40 agonists (soluble CD40L protein or 1 μg/ml coated anti-CD40 antibody), but failed to expand B cells, actually B cells did not proliferate at all, though IgG could be detected in supernantant.
PLEASE help me with expanding a single B cell without feeder cells in vitro.
BTW, is hCD40L-293T better than -3T3 for expanding human memory B cells?
THANK YOU!
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