تولید بهینه سلول های دندریتیک رت از مونوسیت های خون محیطی به صورت In Vitro

Title

The Optimum Generation of Rat Dendritic Cells from Monocytes in Vitro

English Abstract

Introduction: Dendritic cells (DC) are the most important cells in the initiation of immune responses. Because of their superior antigen-presenting capacities, DC may be useful for the initiation of antitumor responses. Current study was done to give rise to the method of differentiation rat monocyte derived DCs in vitro. Materials and Methods: Rats were anesthetized. Then, 6–10 ml heparinized blood was obtained by heart puncture. Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll Hypaque density gradient centrifugation and were seeded in RPMI 1640 supplemented with FBS. After 2 hours of incubation at 37˚C, adherent cells were used for DC generation. These cells were cultured for 7 days in medium containing rat recombinant GM-CSF and IL-4 at a concentration of 50 ng/ml also 20 µg/ml lipopolysaccharides (LPS) was added at day 6 to induce maturation of the MoDC. Results: Morphological analysis of the MoDC showed most of the mature DC were appeared as single cells or loosely adherent aggregates viewed by light microscopy. Conclusion: Since DCs play an essential role in immune responses, a great deal of this research effort has been focused on the practical method for generation and maturation of these cells to obtain MoDC from PBMC.

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