Preliminary studying on dendritic cell culture and its killing effect on gastric carcinoma cell line MKN 45 / 国际肿瘤学杂志

ABSTRACT

Objective To explore the cultivated methods of dendritic cells (DC) and the killing effect of DC stimulated by CpG ODN1826 on gastritic cancer cells MKN45 in vitro. Methods DC was induced from peripheral blood monocytes stimulated by A group ( GM- CSF + IL-4 ), B group ( GM- CSF + IL-4 + TNF- α), C group(nonCpG ODN) and D group( CpG ODN 1826). The surface markers of DC was analyzed via flow cytometry, and the abilities to stimulate proliferation of allogenic lymphocyte by DC and antitumor experiment were detected by MTT assay. Results On day 10, a majority of cells showed typical morphology of DC in D group and B group with visible branching-like and pseudopod-like structures under microscope. The results of flow cytometry showed that there are significantly high expressed co-stimulated molecules such as CD40, CD1a,CD80, CD86 and MHC- Ⅱ in D group compared to other experimental groups ( P ＜ 0.05 ), which dramatically stimulate the proliferation of allogenic lymphocytes and enhance the killing activity of DC on gastric cancer cells. Conclusion This method can acquire relatively high purified DC, and CpG ODN can significantly induce the differentiation and maturation of DC isolated from peripheral blood and enhance the killing activity of DC on MKN45 by stimulating PBMC in vitro.