Comparison of killing effects of human peripheral blood and umbilical cord blood-derived DC-CIK on colon cancer HT29 cells

MU Ying; WANG Weihua; CHEN Shuangfeng; ZHOU Changhui

doi:10.3969/j.issn.1002-266X.2016.40.004

Abstract：Objective To investigate the killing effect of co-cultured umbilical cord blood-derived dendritic cells (DC) and cytokine-induced killer cells (CIK) on colon cancer cells HT29, to explore the feasibility of using umbilical cord blood derived DC-CIK to substitute peripheral blood derived DC-CIK.Methods Umbilical cord blood (50mL) and peripheral blood (50 mL) was collected respectively.The mononuclear cells were isolated by density gradient centrifugation under aseptic conditions, and DC cells were induced and cultured until their mature.The matured DC and CIK cells were mixed for 8 d, the morphological changes of DC-CIK were observed by inverted microscope.The proliferation of CIK/DC-CIK from umbilical cord blood and peripheral blood were evaluated by cell count assay on day 4, 8, 12 and 16, and specif-ic markers of CIK/DC-CIK ( CD3 +, CD3 +CD56 +) were identified by flow cytometric immunophenotype analysis on day 0 and 16, the specific markers of DC (CD40, CD80, and CD86) were tested on day 0 and 7.The DC-CIK cells were co-cultured in different concentration ratio with HT29 cells (DC-CIK∶HT29 =5∶1, 10∶1 and 20∶1) for 24 h, and the antitu-mor rate of DC-CIK on HT29 was detected by CCK-8, and the apoptosis rate of HT29 was investigated by flow cytometry. Results There was no significant difference in the cellular morphology between umbilical cord blood-derived and peripher-al blood-derived DC/DC-CIK before and after culturing, as well as in the specific markers of DC ( CD40, CD80, and CD86).After 16 days culture, the proliferation rate of CD3 +, CD3 +CD56 +cells from umbilical cord blood was increased significantly as compared with that from the peripheral blood.The stronger killing effect and apoptosis-promoting effect on HT29 cells was found in umbilical cord blood-derived DC-CIK as compared with that of the peripheral blood derived DC-CIK (P<0.05).Conclusion The umbilical cord blood-derived DC-CIK can replace peripheral blood-derived DC-CIK as the seed cells for clinical immunotherapy.