Cd3 antibodies

We induced the ex vivo expansion of peripheral blood mononuclear cells (PBMCs) by coating retronectin at a concentration of 25 mg/mL (Takara Bio, Kusatsu, Japan) and CD3 antibodies at a dose of 5 mg/mL (Biolegend, San Diego, CA, USA) onto the wells of 12-well plates (TPP, Trasadingen, Switzerland). The PBMCs were cultured at a concentration of 0.5 × 10⁶ cells/mL in 2 mL of GT-T551 medium (Takara Bio, Japan) supplemented with 300 U/mL IL-2 (Roncoleukin, Biotech, Moscow, Russia) and 0.6% human blood serum of group AB. Subsequently, the plates were incubated in a controlled environment with 5% CO₂ at 37 °C. During the 2nd and 3rd days of cultivation, we performed regular medium exchanges, replacing half of the culture medium volume with a fresh aliquot of IL-2 to sustain cellular proliferation and metabolic activity.

We stimulated the proliferation of peripheral blood mononuclear cells (PBMCs) by immobilizing retronectin at a concentration of 25 mg/mL (Takara Bio, Kusatsu, Japan) and CD3 antibodies at a dose of 5 mg/mL (Biolegend, San Diego, CA, USA) onto the wells of 12-well plates (TPP, Trasadingen, Switzerland). The PBMCs were cultured at a concentration of 0.5 × 10⁶ cells/mL in 2 mL of GT-T551 medium (Takara Bio, Japan) supplemented with 300 U/mL IL-2 (Roncoleukin, Biotech, Moscow, Russia) and 0.6% human blood serum of group AB. The plates were then incubated in a humidified atmosphere with 5% CO₂ at a temperature of 37 °C. During the 2nd and 3rd days of cultivation, we actively replenished half of the culture medium volume with a fresh portion of IL-2.