November 2021 Share
The field of immunological research is continuously expanding. More and more things are being discovered on a cellular and molecular level. Cell separation and enrichment has become an integral part of studying our immune cell behaviors. One of the most significant challenges in the research community has been finding a high throughput T cell enrichment method that can be used to isolate high volumes of viable cells in short amounts of time. As more and more companies begin to develop their own strategies, alternatives are starting to emerge that offer a higher sort value for a more affordable cost—one of which being Akadeum’s buoyancy-activated cell sorting.
T cell enrichment involves the isolation of a specific T cell population from a larger heterogeneous sample. T cell samples are enriched through positive selection, extracting target cells from the sample and leaving undesired cells behind, or negative selection, which involves the removal of all undesired cells, leaving only the target cells behind. T cells are also enriched by the depletion of additional contaminants, such as red blood cells (RBCs).
Human T cell enrichment provides researchers with a viable sample of healthy T cells which they can study as a reference for the human immune system. Experimenting with a sizable population of one specific cell type gives insight into how those cells function in the body.
Human T cell enrichment typically involves isolating specific subsets of T cells from whole blood samples. Blood is extracted directly from an individual then divided into different portions. There are a multitude of methods available to sort the whole blood sample into RBCs, plasma, and white blood cells (WBCs).
Once the cells are divided into groups, different cell separation methods can be used to isolate more specific cell populations. Methods like density-gradient centrifugation and filtration rely on physical forces to divide the sample based on characteristics such as density and size. Other human T cell enrichment methods, such as fluorescence-activated cell sorting (FACS) and magnetic-based cell sorting, use rapid flows of liquid and magnetic fields to isolate target leukocytes from the greater population. These strategies can be harmful to fragile cell populations and result in more than a few dead target cells.
It should be noted that there is a different process for culturing isolated mouse B cells.
Akadeum Life Sciences has developed an innovative technology called buoyancy activated cell sorting (BACS) that uses microbubbles to provide high throughput T cell enrichment. Like FACS and magnetic-based sorting methods, BACS relies on the reaction between surface antigens on target cells and biotinylated antibodies. Small glass bubbles are coated with streptavidin molecules, which form bonds with biotinylated antibodies specifically designed to target a labeled T cell population. Cells are labeled with the biotinylated antibodies, and once the bubbles bind with labeled cells, they quickly and gently lift them to the top of the solution for collection or removal.
Throughput in cell separation is the speed that enrichment can be completed. This can be in terms of sample volume, number of samples, or the number of cells. How many cells can be isolated in one cycle with the method you’ve chosen? What are the capabilities of a single product or kit? Throughput becomes very important when continuously isolating large amounts of cells.
Akadeum’s BACS allows for high throughput human T cell enrichment for many reasons; the first reason is the speed of the workflow. The entire human T cell enrichment kit process takes only 30-45 minutes including setup and cleanup. With one cell separation taking such a short amount of time, it’s easy to run multiple cell separations throughout one day. Other methods might take hours to set up, carry out, and clean after the completion of separation.
The other primary reason BACS enables high throughput is because it requires no additional equipment. Other T cell enrichment methods require expensive and complex equipment to carry out their procedures. FACS requires a flow cytometer, and magnetic-based sorting methods require large magnets and small magnetic beads. Due to these requirements, unless you have plenty of lab space and resources you will likely only be able to run one cell separation experiment at a time.
The BACS microbubble kit can be done directly in the sample container. There is no limit to the number of samples you can sort simultaneously. Each full kit holds roughly enough for 10 separations at 100 million cells each—that’s a projected one billion cells per kit. The ability to concurrently enrich multiple human T cell samples with such a short workflow makes BACS a suitable high throughput human T cell enrichment alternative to more traditional methods.
BACS addresses many of the long-standing issues in the field of cell separation. It’s a quick, simple, and cost-effective method of human T cell enrichment that can benefit both large companies and independent laboratories.
To learn more about how our microbubbles can increase throughput, check out our page on Cell Enrichment for Immunology Workflows.
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