Shown is the immune cell profile of one individual CT26wt tumor which was isolated at day 10 after tumor cell implantation into the mouse.
Primary tumor material is disrupted, erythrocytes removed, and up to 3x106 single cells/well dispensed. The single cells are stained with the marker panel and analyzed by flow cytometry using an LSR Fortessa (Becton Dickinson). The gating strategy is depicted below, starting at the large arrow in the middle.
|Marker||Immune cell population|
|CD45+; CD11b-; CD3e-; B220+||B cells|
|CD45+; CD11b-; CD3e-; CD49b+||NK cells|
|CD45+; CD11b-; CD3e+; CD8a+||Cytotoxic T cells|
|CD45+; CD11b-; CD3e+; CD4+||T helper cells|
|CD45+; CD11b-; CD3e+; CD4+; CD25+; Foxp3+||CD4+ regulatory T cells|
|CD45+; CD11b+; F4/80+; CD206+; MHCII-||M2 macrophages|
|CD45+; CD11b+; F4/80+; CD206-; MHCII+||M1 macrophages|
|CD45+; CD11b+; F4/80-; Ly6G+; Ly6C+||Neutrophils/Granulocytes|
|CD45+; CD11b+; F4/80-; Ly6G-; Ly6C+||MDCS|
|CD45+; CD11b+; F4/80-; Ly6G-; Ly6C-; CD11c+||Dendritic cells|
Clinical data indicate that only a part of patients responds to treatment with immune-modulating drugs. It has hence been recognized as essential to consider the immune response already during preclinical drug development. Flow cytometry is widely used for the analysis of immune cell populations in tumors.
Panel: A comprehensive analysis of numerous immune cell populations in one tumor is a challenge. We use one staining panel that allows for the analysis of all major immune cell populations and comprises live/dead dye and antibodies against CD3, CD4, CD8a, CD45, CD25, CD11b, Ly6C, Ly6G, F4/80, CD11c, MHC class II, CD206, CD335, CD49b, B220, and FoxP3. The panel can be used for the analysis of tumor tissue and other organs.
Method: Primary tumor material was disrupted, erythrocytes removed, and up to 3x106 obtained single cells/well dispensed. The single cells are stained with the all-in-one marker panel and analyzed by flow cytometry using an LSR Fortessa (BectonDickinson). The gating strategy is depicted below, starting at the large arrow in the middle.
Report: All data of the mouse study, including tumor growth and flow cytometry data, will be presented in a comprehensive report, custom-tailored to the client study, and written by a Ph.D. level technical writer. The analysis enables you to draw conclusions directly from the study outcome and use the data for the filing of official documents.
Add-on cytokine quantification: Based on tumor tissue or blood we can perform cytokine and chemokine quantification with our MESO Quickplex SQ120 assay platform. Please inquire for more information.