The components of the CD40-Ligand Multimer Kit were pre-incubated 30 minutes at room temperature. B cells were isolated using CD19 MicroBeads and cultured (StemMACS HSC Expansion Media XF supplemented with 5% AB serum and 2 µL IL-4) at a final density or 0.15×10
6 cells per mL (low density).
On day 7 and 10 cells were harvested, counted, and restimulated. All cells were harvested, counted and analyzed on day 14. With the low density protocol a B cell expansion of up to 15-fold can be achieved.