Study of differentiation from CML into dendritic cell by calcium ionorphore / 中国免疫学杂志

ABSTRACT

Objective:To explore the methods and condition in which calcium ionorphore(CI) induces the CML cells to differentiate into dendritic cells(DCs).Methods:Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30?30~9 L~ -1 when samples were collected,then lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10%FCS(Fetal calf serum),with or without CI(375 ng/ml) and GM-CSF(200 ng/ml) at 37℃,5%CO_2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI.Results:After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80,CD86,CD40,CD86 and HLA-DR.Conclusion:CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.