Exploratory Research of Setting an Optimal Minor-factor Culture System of Cytokine-induced Killer Cells/ Natural Killer Cells / 中山大学学报(医学科学版)

ABSTRACT

[Objective] In order to explore the optimal minor-factor culture system of CIK/NK cells by proliferating CIK/NK cells using single-factor,bi-factor,and tri-factor combinations of IL-2,IL-7,and IL-12.[Methods] Ficoll-Hypaque method was used to separate cord blood mononuclear cells and divide them into 6 groups.Add single-factor,bi-factor,and tri-factor combinations (IL-7;IL-12;IL-7 + IL-12;IL-2 + IL-7;IL-2 + IL-12;IL-2 + IL-7 + IL-12) of IL-2 (80 ng/mL),IL-7 (40 ng/mL),and IL-12 (40 ng/mL) and culture them in total 21 days then harvest the cells.To collect cell suspensions of each group in culture outset and day 21 and to count the proportion of CD3+ CD56+ CIK cells and CD3- CD56+ NK cells with flow cytometry.[Results] The proportion of CIK cells in single-factor culture system of IL-12 was the highest in all groups (30.23 ± 1.18%).The proportion of CIK cells in bi-factor culture system of IL-2 + IL-12 can raised to 18.58 ± 0.68%.The proportion of CIK cells of the two groups above can reach the level of that using traditional multi-factor culture methods.NK cell proportion in IL-12 was 30.23 ± 1.18%,NK cell proportion in IL-2 + IL-7 can also reach to 29.52 ± 0.89%.[Conclusions] It is adoptable to proliferate CIK/NK cells using minor-factor culture system of IL-12,IL-2 + IL-12,or IL-2 + IL-7.