January 2021 Share
Get an inside look at how Akadeum’s products perform cell separation in real-world research that is happening in laboratories across the country. A research group at the University of Pennsylvania used the Mouse B Cell Isolation Kit from Akadeum Life Sciences in their research into the mechanisms that underlie the differentiation of activated B cells into antibody-producing plasma cells.
Using the Akadeum Mouse B Cell Isolation Kit allowed the research group to obtain a B cell purity of greater than 97%. More importantly, the group observed that the isolated cells were healthy, were not activated by the isolation process, and had maintained their ability to be differentiated into plasma cells.
The Akadeum microbubble workflow can be performed directly in the sample container for quick and easy enrichment – no need for additional equipment and consumables like magnets or columns.
Objective: Identify transcriptional programs and other mechanisms that regulate plasma cell differentiation.
Kit Used: Mouse B Cell Isolation Kit from Akadeum Life Sciences
Procedure: A single cell suspension of splenocytes was obtained by mechanically dissociating spleens of 8-week-old C57BL/6 mice and passing the dissociated material through a filter. The resulting sample was enriched for B cells using Akadeum’s Mouse B Cell Isolation Kit. Post-enrichment, B cells were cultured in the presence or absence of CpG and IL-5 for 3-days and plasma cell differentiation was assessed based on induction of CD138.
Key Takeaways: B cell purity of >97% was obtained while physiology and plasma cell differentiation potential were conserved post-enrichment.
Outcome: The sample preparation using Akadeum’s Mouse B Cell Isolation Kit delivered a highly-enriched sample with B cell purity of greater than 97%. Following plasma cell induction, upregulation of CD138 was confirmed in the enriched B cell population, validating post-enrichment function.
Figure 1: Akadeum’s microbubble kit delivered a highly-enriched sample for downstream analysis. Flow plots for CD19+ B cells before enrichment (left), following enrichment with Akadeum’s Mouse B Cell Isolation Kit (middle), and following positive selection with magnetics competitor (right).
Figure 2: B cells were not artificially activated by Akadeum isolation and maintained their capacity for plasma cell differentiation as shown by CD138 induction. Flow plots from enriched CD19+ B cells that were cultured for 3 days in the absence (left) or presence (right) of IL-5 and bacterial CpG.
Our Mouse B Cell Isolation Kit performed exactly as expected, delivering a highly enriched sample for downstream analysis using our protocol that is fast, easy, and gentle on cells. As the research team themselves said, “Generally very easy to use…it was streamlined compared to magnetic bead separation. The ability to get 4 times the number of cells over competitor is absurd.”
If you are involved in cell isolation research, wondering how to culture isolated mouse B cells, or even looking for a human B cell isolation kit, we’d love to hear from you! We are actively seeking new applications for our microbubble technology and would welcome the opportunity to learn more about your research and to discover if our microbubbles could help you in your work. Let’s schedule a time to talk!
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