Product Details
How it Works:
Akadeum’s novel approach to positive selection for 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.
Using washed leukopak material or PBMCs as a starting material, T cells are labeled with biotinylated antibodies targeting CD3 and CD28. BACS™ Selection, Activation & Expansion Microbubbles are mixed into the sample where they bind to the labeled cells and gently float them to the surface. Unwanted cells can be removed from below the microbubble layer, and the positively selected T cells transferred to a culture vessel.
BACS™ are inherently inert and do not impact culture conditions. Over time and with cell division, cells gently settle to the bottom of the plate where they continue to expand until collection.
Additional Information:
- Compatible with the Alerion™ Microbubble Cell Separation System
- Antibody Cocktail (biotinylated): CD3 and CD28
- Storage: 4°C
- Cell Type: T Cells
- Sample Species and Source: Human PBMC or platelet washed Leukopak material
- Cell Separation Method: Positive Selection
- Capacity: 800 x 106 nucleated cells
This Kit Comes With:
- Selection Activation & Expansion Microbubbles
- Selection Activation & Expansion Antibody Cocktail
View the Checklist for Using Akadeum Cell Therapy Kit Products
Key Features
Go from research scale to commercialization with microbubbles. Just use more or less to suit your research needs and keep performance maintained.
Experimental Overview:
T cells were positively selected from PBMC input material at 100 million total cells (5 mL tube), 1 billion total cells (a 150 mL bag), or 5 billion total cells (BioRise™ Consumable and Alerion™ Microbubble Cell Separation System) following the Human T Cell Selection, Activation, and Expansion protocol.
A) Estimated time based on protocol steps (mixing, incubation, separation) for separation completed in each vessel. As cell numbers increase the workflow time does not lengthen.
B) Comparison of cell viability results at day 1 post selection for cells separated in 5 mL tube, 150 mL bag, and BioRise™ Consumable. All protocols resulted in high cell viability.
C) Total cell expansion results comparing the BioRise™ Consumable selected cells to 5mL tube and 150mL bag vessel controls. Five million captured T Cells were seeded for expansion using G-Rex® 6M Well Plates and complete ExCellerate™ media supplemented with 2% human serum. Additionally, 5U/mL IL2, 5ng/mL IL7, and 5ng/mL IL15 was added on days 0, 4, and 7 of culture. No significant differences were observed in growth rates between the different isolation vessels.
The Right Products to Support Your Pipeline
The Human T Cell Selection, Activation and Expansion Kit is ideal for research through process development, with GMP and Clinical Ready versions available to support preclinical to commercialization of cell therapy workflows.
Data
More Activation, Less Exhaustion. Healthier Cells.
Optimize results while minimizing potential cellular stress or damage.
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2 x 108 PBMCs were used as starting material for the Akadeum Human T Cell Selection, Activation, and Expansion kit workflow. Post selection, 5 x 106 T cells/well were seeded and cultured in G-Rex® 6M well plates (Wilson Wolf Manufacturing). Magnetic particles (following the manufacturer’s recommended protocol) were used for comparative T cell selection and were seeded similarly at 5 x 106 per well. Cells for control wells were isolated via Akadeum’s Human T Cell Leukopak Isolation Kit (negative selection) and seeded at 0.5 x 106 per mL in standard 24-well tissue culture plates. Complete ExCellerate™ Media (Bio-Techne), supplemented with 2% human serum (Access Biologicals) was used for all conditions. Additional IL-2, IL-7, and IL-15 cytokines (PeproTech), were added for Akadeum Human T Cell Selection, Activation, and Expansion kit wells while magnetics and control wells were supplemented with IL-2. Harvest timepoints for all conditions were on days 4, 7, and 11.
