In vitro culture of bone marrow-derived dendritic cells of mouse and a study of their function
Wang Hongwei?
Lu Jiangyang
Qin Jiajian
Kang Jiarui
Yang Wen
Abstract：Objective:To establish the method for cultivation and amplification of bone marrow-derived dendritic cells (DCs ) of mice in vitro, in order to provide an experimental base for the DC-targeting therapeutic study.Methods:Bone marrow cells of C57BL/6 mice were obtained and cultured in RPMI1640 containing recombi-nant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF)400 U/ml,interleukin-4 (IL-4)200 U/ml,and 10% fetal bovine serum invitro. On the 4th day,lipopolysaccharide (LPS)was added.Two days later, the morphology and ultrastucture of DCs were observed by phase contrast microscope and transmission electron microscope. The purity and immunophenotype of DCs were verified by flow cytometry on day 6. The contents of cytokines including IL-6,IL-10,monocyte chemotactic protein-1 (MCP-1),interferon-1 (IFN-1),tumor necrosis factor-α(TNF-α),IL-12p70 in supernatant were determined by cytometric bead array.Results:After cultured for 4 days followed by stimulation with LPS for 2 days,nearly 90% of mouse bone marrow cells expressed the charac-teristic DC marker-CD11c,with high level of antigen presentation molecule-major histocompability complex-Ⅱ(MHC-Ⅱ)and costimulatory molecules-CD86 and CD80. The cells possessed the typical morphologic feature of DCs and the function of secreting pro-inflammatory cytokines such as IL-6,IL-10,MCP-1,TNF-αand IL-12p70. Conclusions:Culturing invitro with GM-CSF and IL-4,bone marrow cells of mice can be induced to become DCs with high purity,with their characteristics of morphology,immunophenotype,and immunological functions.
Keywords：Bone marrowDendritic cellsGranulocyte-macrophage colony-stimulating factorInterleukin-4
Publication Date：2014-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 199-203 )