Primary T Cell Growth and Transfection
Primary T cells from three healthy donors were purchased from Astarte Biologics and grown in X-VIVO™ 15 media with Human T Activator CD3/CD28 Dynabeads (Thermo Fisher Scientific #11131D) at a 1:1 ratio of beads to cells. The culture media was also supplemented with IL-2 at 0.5 ng/mL to stimulate T cell growth. Beads were replaced every 7-10 days during passaging, while IL-2 was added every 2-3 days. Transfections of primary T cells followed the protocol that was optimized for the Jurkat T cells (2.75 µL Lipofectamine and 1 µg of pDNA per well in a 24-well plate).