I'm trying to think through the best way to stimulate mouse CAR T cells that target mouse CD19 (1D3 scFv) without incubating the CAR T cells with B cells. I want to do this because a metabolic assay (Seahorse) that I want to run needs to have only stimulated CAR T cells. If there are B cells in the cell culture, then that would skew the results of the assay since I'm only interested in the metabolic profiles of the CAR T cells. Also, sorting the CAR T cells and B cells is not an option because sorting and resting the cells would also alter the metabolic profiles of the CAR T cells.
My initial thoughts is to biotinylate mouse CD19 and then conjugate the CD19-biotin to streptavidin beads. That would allow me to activate my CAR T cells with CD19-conjugated beads similar to using Dynabeads when stimulating conventional T cells.
Is there a better/easier/cheaper way to stimulate my CAR T cells?
You might try using fixed B cells as stimulating agent. I except them to be metabolically inactive. You could verify that including wells with only fixed B cells in your assay.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology
- Enzymes