Culture and Identification of Mouse Bone Marrow-Derived Dendritic Cells and Their Capability to Induce T Lymphocyte Proliferation
Wenguang Wang, Jia li, Kun Wu, Baihetiya Azhati, Mulati Rexiati
Department of Urology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China (mainland)
Med Sci Monit 2016; 22:244-250
The aim of this study was to establish a culture method for mouse dendritic cells (DCs) in vitro and observe their morphology at different growth stages and their ability to induce the proliferation of T lymphocytes.
MATERIAL AND METHODS: Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) were used in combination to induce differentiation of mouse bone marrow (BM) mononucleocytes into DCs. The derived DCs were then assessed for morphology, phenotype, and function.
RESULTS: The mouse BM-derived mononucleocytes had altered cell morphology 3 days after induction by GM-CSF and IL-4 and grew into colonies. Typical dendrites appeared 8 days after induction. Many mature DCs were generated, with typical dendritic morphology observed under scanning electron microscopy. Expression levels of CD11c, a specific marker of BM-derived DCs, and of co-stimulatory molecules such as CD40, CD80, CD86, and MHC-II were elevated in the mature DCs. Furthermore, the mature DCs displayed a strong potency in stimulating the proliferation of syngenic or allogenic T lymphocytes.
CONCLUSIONS: Mouse BM-derived mononucleocytes cultured in vitro can produce a large number of DCs, as well as immature DCs, in high purity. The described in vitro culture method lays a foundation for further investigations of anti-tumor vaccines.
Keywords: Bone Marrow Cells - ultrastructure, Animals, Cell Culture Techniques - methods, Cell Proliferation, Cell Shape, Cells, Cultured, Dendritic Cells - ultrastructure, Flow Cytometry, Lymphocyte Activation - immunology, Lymphocyte Culture Test, Mixed, Mice, Inbred C57BL, T-Lymphocytes - cytology, Time Factors