Description

About the C57BL/6 Mouse

The C57BL/6 mouse is the most commonly-used inbred mouse strain and is employed in numerous research areas, including immunology, genetics, diabetes and obesity, neurobiology, cardiovascular biology, and developmental biology. They are routinely used as the background strain for generation of congenics carrying modified genes and/or transgenes. These mice carry the H-2b haplotype and their immune cells are commonly used in immune-based assays. Further, the C57BL/6 mouse is biased towards a Th1 cell-mediated immune response when challenged appropriately. Together, the C57BL/6 mouse has been extensively used to study numerous scientific questions and their cells (like these C57BL/6 mouse PBMCs) may offer many experimental benefits depending on the parameters of your experiments.

C57BL/6 CD3 Pan T Cells

CD3 pan T cells from C57BL/6 mice consist of both CD4 and CD8 T cells and make up approximately 33% of cells in the spleen of a typical adult animal. All T cells carry the CD3 protein marker and the T cell receptor (TCR), which recognizes foreign antigens presented by major histocompatibility complex molecules on antigen presenting cells, to activate them.

T cells develop in the thymus to become mature CD4 or CD8 T cells and exit the thymus to reside and encounter foreign pathogens in lymphoid tissues.

When activated, CD4 T cells will become T helper subsets to secrete cytokines to boost either the humoral or cell-mediated immune response. In contrast, CD8 T cells, when stimulated, will become cytotoxic T cells that can lyse infected or transformed cells. Together, these two main T cell subsets work to clear infections or diseased cells.

C57BL/6 Mouse Pan T Cell Application Summary

Purified C57BL/6 pan T cells are a good source of cells to study the biology of T cells and their role in the immune system. T cells are responsible for producing cytokines and performing cytotoxicity. Thus, pan T cells can be used for a variety of stimulation-dependent functional experiments to assess cytokine production, proliferation, and cytotoxicity.

With the current trend toward development of immuno-modulatory drugs, purified mouse pan T cells are an ideal source of cells to test T cell response in the preclinical phase of therapeutic molecule development.

C57BL/6 Mouse CD3 Pan T Cell Purification

Spleen Collection and Splenocyte Isolation

Spleens are isolated responsibly from C57BL/6 mice at third party facilities. These facilities are vetted to ensure they operate according to local regulations and laws. Since the facility is local, the tissue is quickly transported to iQ’s facility for processing.

Once at our facility, the splenic tissue is carefully dissociated through a gentle mechanical process to generate a single cell suspension under sterile conditions. The suspension is then carefully passed through a filter to remove any fat and/or connective tissue and subsequently depleted of red blood cells in a manner that is not damaging to the other cells. The purified splenocytes are then used to isolate pan T cells.

Isolation of C57BL/6 Mouse CD3 Pan T cells

To enrich for pan T cells by the negative isolation method, splenocytes were incubated with antibodies against B cells, NK cells, dendritic cells, monocytes, granulocytes, and erythrocytes in a test tube, and subsequently subjected to a magnet. Cells labeled with the antibodies bound to the magnet through the test tube wall, while unlabeled cells, the pan T cells, were decanted into a fresh tube to obtain the enriched population.

Contact us for more information about purchasing our magnetically isolated pan T cells.

Purity

A small aliquot of cells was tested for post-sort purity by flow cytometry analysis. Purity of pan T cells, as defined by CD3 expression, was ≥ 90%.

Cryopreservation and storage

Our C57BL/6 mouse pan T cells are cryopreserved carefully using iQ Biosciences’ cryopreservation protocol that ensures high viability (typically > 70%) after thawing.

Cells should be stored at < -120°C once they are received, such as within a liquid nitrogen tank (vapor phase).