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Excerpt
Aims: Since the first report by Klinkert and Bowers, it has been well established that specialized antigen presenting cells (APC), i.e., dendritic cells (DC) are obtained from culturing bone marrow cells (BMC) and that DC are found to bear distinct phenotypes and function from macrophage (Mφ). Nevertheless, dendritic cells are generally difficult to obtain in a substantially large number for many experimental purposes due to the paucity in the peripheral lymphoid tissues. In this regard although it has been reported that granulocyte macrophage colony-stimulating factor (GM-CSF) is one of the potential candidates that is able to support both growth and differentiation from DC precursor and/or its progeny in mouse BMC, it has been reported that co-stimulatory factors such as tumor necrosis factor (TNF-α) and/or IL-4 are effective to enhance human and rat DC induction from BMC culture. Inasmuch as GM-CSF appears to be not a solely factor for DC growth and differentiation, it is important to determine what is a crucial cytokine to induce cell growth and differentiation DC from its BMC progeny. Furthermore heterogeneity of their cell origin, differential function and growth requirements have been reported. Thus, it is important to identify a specific factor and to establish a general principle to induce growth and differentiation of DC from their progeny.
Methods: We examined any beneficial effects of various recombinant cytokines and/or cytokine combinations on inducing the growth and/or differentiation of DC precursors from rat BMC. In particular, employing well-known hematopoietic factors, i.e., c-kit ligand and Flt3/Flk2 ligand, we also attempt to determine whether and to what extent these factors increase the frequency of DC precursor(s) from hematopoietic stem cells to obtain a practical amount of mature DC from BMC culture systems.
Results and Conclusions: Our study demonstrated that unlike mouse systems, we were not able to obtain any evidence to support the idea that GM-CSF per se as a growth factor for DC progeny in BMC regardless of the cytokine resources. However, we obtained the most efficient outgrowth of the DC precursors only with cytokine combinations, Flt3/Flk2 ligand and IL-6, not with c-kit ligand and IL-6. Thus fully mature DC were obtained as much as 100% of cell yields following a differentiation of the DC precursors in concert with additional cytokines of such as GM-CSF, TNF-α, and IL-4.
