maryam golara

Dendritic cells (DCs) are a group of bone marrow-derived cells that play a crucial role in innate and acquired immune responses. Bone marrow-derived dendritic cells (BMDC) are used in many studies, so the efficiency and purity of the differentiated cells are essential. This study aimed to investigate the effect of several parameters, including the age of mice, cell culture medium, and swirling of the culture plate, to increase the efficiency of the induced cells, considering the standard protocols. Bone marrow-derived dendritic cells were induced from both juvenile and adult mice bone marrow cells. Then, the purity of CD11c+ cells was compared between juvenile mice BMDCs and adult mice BMDCs. Cells were cultured in an enriched and non-enriched medium, and some wells were swirled when changing the medium on the 3rd day. Then the effect of enriched medium and swirling before medium replacement were evaluated based on the expression of the CD11c marker. The efficiency of DCs differentiation (CD11c+ cells) was higher when juvenile mouse bone marrow precursors were used compared to adult mice; using the enriched media with supplements and swirling the well before media replacement significantly affected the purity of immature CD11c+ cells. Due to our results, using juvenile mice, an enriched culture medium, and physical removal of granulocyte cells could significantly improve the purity and efficiency of CD11c+ cells. Therefore, considering these three items in the production protocol of these cells can probably reduce the use of lymphocyte-removing antibodies and purification methods.

  Asthma is a reversible chronic inflammatory disease of airways that the effects of inflammatory cytokines in its severity has been proven. This study aimed to compare interleukin 17 (IL17) and interleukin 22 (IL22), two effective cytokines in asthma, children with and without asthma. In this case-control study, 15 children with asthma and 15 healthy children were included. Peripheral blood mononuclear cells (PBMCs) were isolated from 5 cc of heparinized blood samples, and cultured under stimulation with anti-CD3 and anti-CD28 for 72 hours. Percentage of CD4+IL17+ and CD4+IL22+T cells was assessed by flow cytometry method. Secretion of IL17 and IL22 cytokines was measured using enzyme-linked immunosorbent assay (ELIZA) technique. There was no significant difference between cases and controls in levels of secreted IL17 and IL22 in supernatant of cultured cells. Moreover, there was no significant difference between patients and healthy group in terms of CD4+IL17+ (P = 0.30) and CD4+IL22+ (P = 0.41) T cells in PBMCs; although the median percentage of IL17+IL22+ T cells in CD4+ T-cell subset had an elevated level in patients group but the difference was not significant (P = 0.56). In studied community containing children with relative- and completely-controlled asthma, there were no significant differences with healthy children in IL17 and IL22 levels, as well as CD4+IL17+ and CD4+IL22+ T cells in PBMCs. Although the median percentage of CD4+IL17+IL22+* T cells had not a significant elevated level in patients group, more studies with larger sample sizes could be helpful.