This study aims to develop a method to efficiently and stably induce the differentiation and expansion of human myeloid-derived suppressor cells(MDSC)in vitro and provide new experimental technology for the study of the function and mechanism of human MDSC.For this purpose,PBMC of healthy volunteers were isolated with Ficoll density gradient centrifu-gation.Next,monocytes were separated by anchoring technique.PBMC or monocytes were stimulated with GM-CSF and IL-6(10,20,40,80 ng/mL each)for 4,7,14,and 21 days before co-culturing with PBMC for 3 days.The proliferation of CD33 +MDSC and T cells and the proportion of CD3+ IFN-y+ T cells were detected by FACS.The results showed that the PBMC or monocytes were induced to differentiate to less CD33+ MDSC by conventional methods.However,by improved methods,the monocytes cultured with GM-CSF and IL-6(each 80 ng/mL)for 14 days(refreshing the culture solution every 4-5 days)were induced to differentiate more efficiently into a higher proportion of CD33+ MDSC,which showed significant inhibition of T cell proliferation and IFN-y secretion.This study suggests that with the improved method,the monocytes are efficiently induced to differentiate into CD33+ MDSC,which has significant suppressive function.
关键词
人髓源抑制性细胞/体外分化/实验技术
Key words
human myeloid-derived suppressor cell/differentiation in vitro/experimental technology
维普
万方数据