Establishment of a method for in vitro amplification of murine bone marrow dendritic cells

Objective: To establish a method of expanding dendritic cells from mice bone marrow in vitro. Methods: Cells isolated from mice bone marrow were cultured in GM-CSF and IL-4. Morphological change was observed by optical phase contrast microscopy, and T cell proliferation was detected by 3H-TdR incorporting into assay, and surface molecular by FACS, and cytokines were assayed by ELISA. Results: A large number of morphologically typical DCs were observed after the culturing for 8 days, and DCs had potent ability of stimulating auto and allogenic T cell proliferation. FACS analysis showed that the expanded DCs could express molecular of IaK b,CD11c, CD80, CD86 and ICAM-1. LPS promoted DCs to be mature and secreted higher levels of IL-12(p70), IL-6, TNF-α and, IL-1β. Conclusion: A large number of mature DCs can be generated by in vitro culture of mice bone marrow cells.