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Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice
Published on: July 1, 2022
[β-Glucan promotes the maturation and migration of bone marrow-derived dendritic cells]
Dongqin Xu1, Xiaohang Zhang1, Yong Wang1
1Medical Research Center, Changzhou Second People's Hospital, Nanjing Medical University, Changzhou 213000, China.
Objective:
To investigate the effects of β-glucan on the maturation and migration of bone marrow-derived dendritic cells (BMDCs).
Methods:
BMDCs were isolated from mouse bone marrow cells in vitro and induced by β-glucan for maturation. The expressions of cell surface markers were detected by flow cytometry (FCM). The cytokines (IL-6, IL-12p40, tumor necrosis factor α) in the supernatants were measured by ELISA, and the expressions of intracellular CC chemokine receptor 1 (CCR1), CCR2, CCR5, CCR7 were determined by real-time quantitative PCR. Furthermore, the chemotactic response to CC chemokine ligand 19 (CCLl9) and CCL21, i.e. CCR7-1igands, was measured by Transwell(TM) migration assay. Moreover, the number of migrated cells in the draining lymph nodes was analyzed by FCM.
Results:
Compared with the control group, the expressions of co-stimulation molecules (MHC II, CD40, CD80, CD86) on BMDCs were up-regulated in the presence of β-glucan. Furthermore, β-glucan could prompt BMDCs to secret high levels of IL-6, TNF-α, IL-12 p40 and increase the production of CCR7 mRNA. After β-glucan treatment, BMDCs were more sensitive to CCL19/CCL21. The number of BMDCs migrated from subcutaneous injection site into the draining lymph nodes significantly increased in β-glucan group.
Conclusion:
β-glucan can promote the maturation of BMDCs and enhance the migration ability of BMDCs in vitro and in vivo.
