| Objective:To explore an efficient method for inducing and expanding CD56+ lymphocytes from mononuclear cells of human peripheral blood(PBMCs) and bone marrow in vitro.Method:Mononuclear cells from human peripheral blood(PBMCs) and bone marrow were cultured in stem cell growth medium(SCGM) supplemented with IL-2/anti-IL-2 complex,IL-2,anti-IL-2 mAb and IL-2+IL-15 respectively to induce and expand CD56+ lymphocytes,the immune phenotypes of cells from blood were analyzed by Flow Cytometry(FCM) on the 7th and 10th day after incubation separately,while those from bone marrow were analyzed on the 7th and 14th day.The cytotoxicity of cells from peripheral blood and bone marrow to K562 targets was detected by MTT assay after culturing for 10 days and 14 days separately.Results:The CD56+ cells from peripheral blood cultured with IL-2/anti-IL-2 complex got the best expansion and expanded to 24.67±3.14 folds in 7 days and 31.63±5.01 folds in 10 days after incubation;Among them,the expanded folds for CD3+CD56+ and CD3-CD56+ were(110.93±19.10),(7.8±1.17) after culturing for 7 days and(157.60±46.31),(12.03±1.64) after 10 days respectively;while those from bone marrow expanded to11.08±2.08 folds at the 7th day and 21.72±2.19 folds at the 14th day, of which the expanded folds for CD3+CD56+ and CD3-CD56+ were(30.30±5.14),(6.05±1.05) after culturing for 7 days and(60.18±6.68),(10.75±1.51) after 14 days respectively.At 10:1 of effecter/target ratio,the cytotoxicity of cells from peripheral blood and bone marrow cultured with IL-2/anti-IL-2 complex to K562 cells increased up to(83.46±1.56)%and(74.94±2.28)%respectively,significantly higher than the other control groups.Conclusions:CD56+ lymphocytes could be expanded effectively from PBMC and bone marrow using SCGM under the stimulation and induction of IL-2/anti-IL-2 complex.This work provided a novel and effective method of expanding human CD56+ lymphocytes in vitro for the adoptive immunotherapy of malignant tumors. |
