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).