Akadeum’s Human T Cell Activation & Expansion Kit generates large numbers of highly activated, less exhausted cells, enabling better science and more effective cell therapies. Using BACS™ microbubble technology, the Human T Cell Activation & Expansion Kit was designed to isolate highly pure populations of T cells and activate them for use in cell therapy workflows. This highly scalable isolation, activation, and expansion process yields the large numbers of highly activated and transducable cells needed to manufacture potent cell therapies. In addition, the gentleness of flotation-based culture results in fewer exhausted cells than common activation methods. Akadeum’s novel approach to CAR-T cell production provides improvements over current industry standard methods, and the innate scalability of BACS™ microbubbles allows for rapid optimization of cell therapy workflows.
Mechanism of Action: A process that combines selection, activation, and expansion into a single workflow powered by BACS microbubbles is presented in this study. Due to the natural buoyancy of the microbubbles, activation and expansion take place at the top surface of the culture vessel. As the stimulated T cells divide, daughter cells dissociate from the microbubbles and settle at the bottom surface of the culture vessel where they are no longer in contact with the activation signals. This separation from the activation signals prevents overstimulation and results in a 2-fold lower expression of exhaustion markers relative to the industry-standard activation methods – only 25-30% PD-1+ T cell when microbubbles are used compared to over 90% when iron particle-based methods are used. The Human T Cell Activation & Expansion kit reduces the expression of exhaustion markers without compromising T cell activation as evidenced by the rapid upregulation of CD25 and other standard activation markers. Additionally, buoyant activation using microbubbles yield enhanced effector T cell populations and industry standard expansion kinetics.
How it Works: Starting from leukopak material or PBMCs, T cells are labeled with biotinylated antibodies targeting CD3 and CD28. BACS™ Activation & Expansion Microbubbles are mixed into the sample where they bind to the labeled cells and gently float them to the surface. Unwanted cells are removed from below the microbubble later, and the positively selected T cells are transferred to a culture vessel. As the cells divide daughter cells gently settle to the bottom of the plate, ready for collection.
- Storage: 4°C
- Cell Type: T Cells
- Sample Species and Source: Human PBMC or Leukopak material
- Cell Separation Method: Positive Selection
- Capacity: 800 x 106 nucleated cells
This kit comes with:
- BACS™ Activation & Expansion Microbubbles
- CD3/CD28 Activation & Expansion Antibody Cocktail
Recommended for Best Results
- 20 rpm tube rotator for mixing (e.g., Thermo Scientific cat#: 88881002)
- Centrifuge (swinging bucket rotor strongly preferred)
- 9-inch sterile Pasteur Pipets
- 5 mL / 50 mL tubes
- Culture plates or bags