Using Akadeum’smicrobubble technology, the Human T Cell Selection, Activation, and Expansion Kit was designed to isolate highly pure populations of T cells and activate them for use in cell therapy workflows and assays. T cells are isolated, selected, activated, and expanded using Akadeum’s activation and expansion microbubbles in combination with an antibody cocktail that targets CD3 and CD28.
This immensely scalable process yields large quantities of highly activated and transducable cells needed to manufacture potent cell therapies. In addition, the gentle flotation-based culture results in fewer exhausted cells than traditional activation methods.
Other benefits include:
Scalable to fit your cell therapy workflow needs
Closed-system in-bag workflows available
Reduced cellular exhaustion during T cell activation and expansion
Increased numbers of highly activated effector T cell populations
Industry-standard T cell expansion kinetics
Request a Human T Cell Selection, Activation, and Expansion Kit Quote
Akadeum’s Human T Cell Selection, Activation, and Expansion Kit efficiently produces large quantities of activated T cells with reduced exhaustion levels. The T cell kit utilizes microbubble technology to extract and activate T cells with high purity.
This effective and scalable research-use method results in a substantial number of activated and transducible T cells, crucial for creating powerful cell therapy treatments. Moreover, the T cell kit’s flotation-based culture technique leads to a lower incidence of cell exhaustion compared to traditional human T cell activation methods.
Akadeum’s novel approach to CAR-T cell production provides improvements over current industry standard methods and reagents. The innate scalability of microbubbles allows for rapid optimization of immunotherapy workflows and assay integration.
Mechanism of Action
The Human T Cell Selection, Activation, and Expansion Kit combines selection, activation, and expansion into a single workflow powered by microbubbles. Due to the microbubbles’ natural buoyancy, activation and cell 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 two-fold lower expression of exhaustion markers relative to the industry-standard activation methods.
For example, only 25-30% of T lymphocytes express the PD-1 exhaustion marker when microbubbles are used compared to over 90% when magnetic particle-based methods are used (see Data tab). Cells cultured using Akadeum’s microbubble platforms will be primed for reactivation when introduced back into the body.
The kit reduces the expression of exhaustion markers without compromising activation, as evidenced by the rapid upregulation of CD25 and other standard activation markers.
How it Works
Starting from leukopak material or peripheral blood mononuclear cells (PBMCs), T cells are labeled with biotinylated antibodies targeting CD3 and CD28, mimicking antigen presenting cells in the body. Selection, Activation, and Expansion Microbubbles are mixed into the sample where they bind to the labeled cells and gently float them to the surface for proliferation.
Unwanted cells are then removed from below the microbubble layer, and the positively selected T cells are counted using flow cytometry and transferred to a culture vessel. As the activated cells divide, daughter cells gently settle to the bottom of the plate, ready for collection.
Additional Information:
Storage: 4°C
Cell Type: T Cells
Sample Species and Source: Human PBMC
Cell Separation Method: Positive Selection
Capacity: 800 x 106 nucleated cells
What’s Inside the Kit:
Akadeum’s Selection, Activation, and Expansion Microbubbles
CD3/CD28 activation and expansion antibody cocktail
Akadeum’s Human T Cell Selection, Activation, and Expansion Kit generates large numbers of highly activated, less exhausted cells, enabling better science and more effective cell therapies. Using Akadeum’s microbubble technology, the Human T Cell Selection, Activation, and Expansion Kit was designed to isolate highly pure populations of T cells and activate them for use in cell therapy workflows. Akadeum’s novel approach to CAR-T cell production provides improvements over current industry standard methods, and the innate scalability of microbubbles allows for rapid optimization of cell therapy workflows.
Elegant in Simplicity, Powerful in Application
Akadeum’s microbubble technology harnesses the power of flotation to streamline your immunotherapy workflow. Our novel technology couples performance, ease, and quality, bringing a new dimension to your cell culture.
What Are the Advantages of Akadeum’s Microbubble Workflow?
Minimal expression of exhaustion markers:
Microbubble-enabled cultures result in significantly less expression of the exhaustion markers PD-1, TIM-3, and LAG-3 compared to magnetics.
No compromise to expansion kinetics:
The Akadeum Human T Cell Selection, Activation, and Expansion Kit yields greater than 20-fold expansion over 10 days of culture.
Strong effector cytokine profile:
Microbubble activation results in strong expression of effector cytokines.
Efficient for transduction, gentle on cells:
T cells activated via microbubbles are highly transducible while remaining healthier and less stressed at the same time.
Looking for Answers?
At Akadeum Life Sciences, our team is committed to your success. We thrive on problem-solving and dedicate ourselves to helping researchers obtain exceptional results.
If you’re interested in leveraging our microbubble platform and related products to overcome existing hurdles in your cell therapy workflow, we want to hear from you! You can speak directly with an Akadeum scientist available to help you with an optimized workflow. Schedule a call today; we’d love to start a conversation!
Data
In the experimental results below, T cells were isolated from PBMCs using the Human T Cell Selection, Activation, and Expansion Kit. The purified microbubble-bound T cells were subsequently cultured for 13 days. Cell quantity fold-change and percent frequency of programmed cell death protein 1 (PD-1) expressing cells were determined in vitro at the indicated times.
Figure 1: Quantities of T cells isolated from PBMCs were cultured for 13 days under either control conditions or with Akadeum’s Human T Cell Selection, Activation, and Expansion. T cells cultured with Akadeum’s kit had a 20-fold increase in cell count after 10 days of culture and over 50-fold after 13 days compared to control T cell expansion cultures.
Figure 2: PD-1 is a known exhaustion marker expressed by effector lymphocytes indicating decreased viability and functionality. Microbubble-enabled cultures resulted in a significantly smaller percentage of effector T cells expressing PD-1 at each culture day measured as compared to those cultured using magnetic bead-based activation and expansion methods.
Resources
Follow along with our expert-in-residence, Jason Ellis, PhD, as he demonstrates the protocol for using Akadeum’s Human T Cell Selection, Activation, and Expansion Kit with PBMCs and apheresis materials.
