Study On Generation Of Dendritic Cells From Chronic Myelocytic Leukemia Cells Induced By Calcium Ionophore

Objective: To investigate the effects of Calcium ionophore(CI) on the induction of chronic myelogenous leukemia cells into dendritic cells (CML-DCs). To explore a rapid, simple and efficient way to generate dendritic cells from chronic myelogenous leukemia cells.Methods: Bone marrow mononuclearcells were isolated from chronic myelogenous leukemia cells by density gradient centrifugation using Ficoll-Paque. The cells were cultured with Calcium ionophore (100, 200, 300, 500ng/ml) or cytokines for 9 days. The cytokines were combined with GM-CSF(1×10~3U/ml),IL-4(1×10~3U/ml) and TNF-α(200U/ml). The morphology of DCs was identified by inverted microscope and scanning electron microscope. The survival cells were counted at the day of 0, 1, 3, 5, 7, 9 and calculated the viability by Trypan Blue exclusion. The phenotype (CD83, CD80, CD86) of CML-DCs was analysed by flow cytometry. The capability of stimulating allo-lymphocyte or auto-lymphocyte proliferation with CML-DCs was tested with mixed leukocyte reaction using MTT assay. The migratory capacity of CML-DCs was evaluated in a Transwell system.Results: Bone marrow monocytes from CML patients, which were cultured in Calcium ionophore or in cytokine medium show typical DC morphology. Larger numbers of the cells were ruffled, and stretched out spinous processes that varying length and number, resulting in a variety of cell shapes. After 24 hours of cluture,the forms of CI cultured CML cells changed, and had a typical DC morphology after 72 hours. The CK cultured cells stretched out spinous processes after 5...