Enhanced Enrichment of iNKT cells from Mouse Splenocyte Preps Following Gentle and Rapid Depletion of Erythrocytes and B220+ B Cells with Akadeum's BACS Microbubbles

Biological samples are incredibly diverse and complex, and an initial cleanup step is essential to reduce sample heterogeneity prior to downstream applications. This is especially important when dealing with cell types of low abundance like Invariant natural killer T (iNKT) cells. iNKT cells are a subset of unconventional T cells that recognize lipid-based antigens and have been shown to have a potent anti-tumor response when stimulated appropriately. This has spurred interest in identification of iNKT cell activators as potential adjuncts to immunotherapy regimens. As iNKT cells represent less than 1% of splenocytes or cells in peripheral blood, isolating enough high-quality iNKT cells for the studies necessary for advancing our understanding of their potential can be difficult and time consuming. 

To address the problems created by sample heterogeneity, Akadeum's microbubble platform was leveraged for depletion of both erythrocytes and B220+ B cells from mouse spleen preps as a means for enriching low abundance T and iNKT cell populations. The Mouse RBC and B cell depletion microbubbles removed over 95% of the RBCs and over 93% of the B cells, and a marked decrease in the amount of residual cell debris was observed following the Akadeum depletion protocol as compared to traditional ACK lysis buffer treatment. This depletion of RBCs and B cells coupled with a reduction in cellular debris resulted in a sample significantly enriched for T and iNKT cells. These isolated cells were of the highest quality and were ultimately used for in vivo proliferation assays. 

Sequential application of these simple 10-minute sample clean up protocols saved time and resources, improved sample quality, and led to more meaningful results. This work clearly demonstrates the value of Akadeum’s easy, gentle, and effective cell isolation platform.

Akadeum's Microbubbles represent the next generation of cell separation technology, offering greater than 15x enrichment of iNKT cells.

Download the poster to learn more:

  • Rapid – 3 steps and only 10 minutes to complete each depletion
  • Adaptable – Sequential isolations facilitate customizable cell enrichment protocols
  • Effective – Over 95% of TER119+ and more than 93% of B220+ cells removed
  • Gentle – Improved viability compared to harsh lysis buffers
  • Cleaner samples – Significantly less dead cells and debris than lysis
  • Enables analysis of rare cell types – Greater than 15x enrichment of iNKT cells enabled transfer of enough iNKT cells to be detected in recipient mice