Figure 1. Expansion and Maturation of Human B Cells with ImmunoCult™-ACF Human B Cell Expansion Supplement

B cells isolated from human peripheral blood mononuclear cells (PBMCs) (Leukopak) using EasySep™ Human Pan-B Cell Enrichment Kit were seeded at 1 x 10^5 cells/well in 24-well tissue culture plates with ImmunoCult™-ACF Human B Cell Expansion Supplement. ImmunoCult™-XF B Cell Base Medium was used as a base medium. The cells were passaged every 3 - 4 days. (A) Fold expansion of viable cells is shown for n = 12 donors, with bars representing the mean and 95% confidence level (range 38- to 1,190-fold at day 14 ± 1 day). (B) Expression of CD138 and CD20 was analyzed by flow cytometry at each timepoint (data represent % positive viable cells; mean ± 1 SD). The observed changes indicate maturation of B cells to plasma cells/blasts.

Figure 2. Light Microscopy Image of Cultured Human B Cells

B cells isolated from human PBMCs (Leukopak) using EasySep™ Human Pan-B Cell Enrichment Kit were seeded at 1 x 10^5 cells/well in a 24-well tissue culture plate with ImmunoCult™-ACF Human B Cell Expansion Supplement. ImmunoCult™-XF B Cell Base Medium was used as a base medium. The cells were passaged on day 4 after seeding and imaged at 40x magnification on day 6.

Figure 3. Expansion and Viability of Human Memory B Cells Cultured with ImmunoCult™-ACF Human B Cell Expansion Supplement

Memory B cells were isolated from human PBMCs (Leukopak) using EasySep™ Human Memory B Cell Isolation Kit and were seeded at 0.5 x 10^5 cells/well in 48-well tissue culture plates with ImmunoCult™-ACF Human B Cell Expansion Supplement. ImmunoCult™-XF B Cell Base Medium was used as a base medium. The cells were passaged every 2 - 4 days and the fold expansion of viable cells (A) and cell viability (B) were calculated at each time point. Data represent the mean ± 1 SD of triplicate cultures for the same donor.

Figure 4. Expansion and Viability of Human Naive B Cells Cultured with ImmunoCult™-ACF Human B Cell Expansion Supplement

Naive B cells were isolated from human PBMCs (Leukopak) using EasySep™ Human Memory B Cell Isolation Kit and were seeded at 0.5 x 10^5 cells/well in 48-well tissue culture plates with ImmunoCult™-ACF Human B Cell Expansion Supplement. ImmunoCult™-XF B Cell Base Medium was used as a base medium. The cells were passaged every 2 - 4 days and the fold expansion of viable cells (A) and cell viability (B) were calculated at each time point. Data represent the mean ± 1 SD of triplicate cultures for the same donor.

Figure 5. Expansion of Human Pan-B Cells Cultured with Various Base Media Supplemented with ImmunoCult™-ACF Human B Cell Expansion Supplement or 10% FBS

Pan-B cells were isolated from human PBMCs (Leukopak) using EasySep™ Human Pan-B Cell Enrichment Kit and seeded at 1 x 10^5 cells/well in 24-well tissue culture plates with various base media supplemented with either ImmunoCult™-ACF Human B Cell Expansion Supplement or 10% FBS. The cells were passaged every 3 - 4 days and the fold expansion of viable cells was calculated at each time point. Data represent the mean + 1SD for n = 3 donors (each culture condition was performed in triplicate).