Ex vivo expanation T CD4+ and T CD8+ isolated from human peripheral blood mononuclear cells

Abstract

Immunotherapy is currently an attractive method in the treatment of cancer. Immune cells are isolated, activated, ex vivo proliferated, functionally manipulated, and then given back to the patient. In this study, CD4⁺ and CD8⁺ T cells from human Peripheral Blood Mononuclear Cells (PBMCs) were isolated, ex vivo activated and cultured under the conditions of using coated magnetic beads CD3/CD28 at the ratio of 01 bead: 01 cell supplemented with 30 U/mL IL-2 in the culture medium. The number of cells was counted every day by staining with Trypan Blue.The percentage of CD3⁺CD4⁺ and CD3⁺CD8⁺ cell populations was confirmed by flow cytometry. After 10 days, the CD4⁺ T cell could proliferate strongly; the total cell number is approximately 6.5 x 10⁷ cells, increasing more than 30 times compared with the cell number on the first day. For CD8+ T cells, there was also a slight increase in cell number, reaching 3 x 10⁶ cells. Flow cytometry results showed that the CD4+ T cell population still maintained a CD3+/CD4+ population of more than 90% at day 04. In contrast, the CD3+/CD8+ ratio in the CD8+ population was only 40% even though the total cell number increased in this population. Taken together, these data indicated that we were able to isolate and culture ex vivo CD4+ and CD8+ T cells.