Effects Of Immediate TNF-A Exposure On Phenotype And Function Of Dendritic Cells Derived From Cord Blood Mono Nuclear Cells

Document Type : Original Article




Introduction: Effects of immediate TNF-α exposure on phenotype and function of dendritic cells (DCs) derived from cord blood mono nuclear cells
Materials and Methods: Umbilical cord blood MNCs were isolated from healthy mothers and were divided into TNF(+( and TNF (-) groups. Both were cultured using SCF, Flt3L, GM-CSF and IL-4. But, three ng/ml of TNF-α was first added in the culture of TNF(+( group. All cells were cultured for 14 days and matured with TNF-α or LPS for additional four days. Light microscopic and flowcytometric analyses were performed on days 0, 7, and 14 of both cultures. MLR and cytokines assays were used to characterize the function of immature and mature DCs.
Results: Co-culture of cord blood monocytes and hematopoietic stem cells led to the production of DCs with a characteristic veiled appearance and were consistent with a DC panel of surface markers. However, immediate exposure to TNF-α enhanced the survival of culturing cells in the first week of culture and produced mature DCs with higher maturation markers and IL-12 production. Addition of TNF-α as a maturation marker led to the production of matured DCs and also certain immature and hematopoietic stem cells with higher level of IL-10 production.
Conclusion: This study developed a simple, easy and cost effective way to generate DCs from non fractionating mononuclear cells. It seems that primitive DCs and monocytes in the MNCs are contented in the presence of TNF. This will lead different hematopoitic stem cells to myeloid pathway and results in DCs.