This SOP outlines the standard method for thawing and culturing of human CAR-T cells only and may not be valid for other species or cell types.
In this Protocol we:
- Day 1: Cryopreserved CAR-T cells are thawed and cultured overnight in the presence of IL-2. Cryopreserved APC target cell lines are thawed, cultured and passaged several days prior to beginning the experiment.
- Day 2: Enrichment, Staining and Antigen-Stimulation of CD4+ and/or CD8+ T cells for 20* hours.
- Day 1: Loading of CAR-T cells onto IsoCode chip*.