Establishment of Optimal Culturing Method and Biological Activity Analysis of Chicken Bone Marrow Dendritic Cells using Chi-rGM-CSF

Denderitic cells (DCs) play a central role in immune regulation and antigen presentation in animals, but quantitative description of chicken DCs culturing method employing rGM-CSF and optimal biological response is still lacking. Therefore, we designed the study to obtain the chicken recombinant granulocyte-macrophage colony-stimulating factor (chi-rGM-CSF), expressed in E. coli BL21 and established a standard concentration unit of chi-rGM-CSF to generate optimal biological respondent denderitic cells from chicken bone marrow. DCs were divided into two groups, (A and B) each was employed two level of chi-rGM-CSF (10 and 20 ng/mL) respectively, co-cultured with interleukin-4 (IL-4, 10 ng/mL), and treated with lipopolysachride (LPS, 1 mu g/mL). The results showed that chi-rGM-CSF was effective to induce avian DCs growth. However, no significant difference was observed in cell proliferation, morphology and phenotype between group A and B, but the mRNA expression levels of surface marker genes, MHC-II, CD40, CD80, CD83 and CD86 were up regulated (<0.05) after LPS stimulation DCs in group B showed significantly higher response to LPS with increased mRNA expression levels of pro-inflammatory cytokines (INF gamma, IL-1 beta, IL-2 and IL-6) and anti-inflammatory cytokine (IL-10) as compared with group A. Therefore, it could be concluded that E. coli expressed rGM-CSF with concentration unit (20 ng/mL) is more effective to obtain optimal culturing and biological response of avian BMDCs. (C) 2013 Friends Science Publishers