Figure 1. Protocol Diagram for Culturing Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured in complete Mouse B Cell Expansion Medium as described in Directions For Use (steps 1 - 7). B Cells were harvested on day 9 for analysis. Cells can also be harvested at earlier time points depending on different applications.

Figure 2. Expansion of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured as described in Figure 1. Fold expansion of viable cells is shown with bar graphs representing the mean ± SEM (n = 8). B cells expanded 176.9 ± 29.8-fold after 9 days of culture.

Figure 3. Maturation of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured as described in Figure 1. Following staining using the protocol by Pracht et al. (Eur J Immunol, 2017), the expression of A) B220 and CD138 and B) TACI (CD267) and CD86 were analyzed by flow cytometry at several time points (data represents mean ± SEM, n = 8). An increase in CD86 cell surface expression indicates B cell activation; a decrease in B220 and an increase in CD138 and TACI cell surface expression indicate maturation of B cells to plasmablasts or plasma cells.

Figure 4. Robust Growth of Mouse B Cells when Cultured with ImmunoCult™ Mouse B Cell Expansion Kit

B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured as described in Figure 1. Cells were imaged at A) 10X magnification on day 0, B) 10X magnification on day 3, C) 40X magnification on day 6, and D) 40X magnification on day 9.