Main Article Content
Bone marrow, dendritic cells, primary culture
Bone marrow derived Dendritic cells (BMDCs) are routinely employed in cell based assays to evaluate immunomodulatory and anti-inflammatory activities. Hence, simplified, stepwise, defined and standardized methods are required for isolation of bone marrow cells from mice, propagating them in presence of growth factors and obtaining high and reproducible yields of BMDCs. Here, we describe a detailed, stepwise protocol with pictorial representation to isolate bone marrow from mouse femur and development of dendritic cells. Mouse bone marrow cells are cultured in presence of granulocyte-macrophage colony stimulating factor (GM-CSF) for 6 days to generate BMDCs.
 Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392:245-252.
 Stockwin LH, McGonagle D, Martin IG, Blair GE (2000) Dendritic cells: immunological sentinels with a central role in health and disease. Immunol Cell Biol 78:91-102.
 Schlichting CL, Schareck WD, Nickel T, Weis M (2005) Dendritic cells as pharmacological targets for the generation of regulatory immunosuppressive effectors. New implications for allo-transplantation. Curr Med Chem 12:1921-1930.
 Abe M, Thomson AW (2003) Influence of immunosuppressive drugs on dendritic cells. Transpl Immunol 11:357-365.
 Liu Q, Shu X, Sun A, Sun Q, Zhang C, et al. (2008) Plant-derived small molecule albaconol suppresses LPS-triggered proinflammatory cytokine production and antigen presentation of dendritic cells by impairing NF-kappaB activation. Int Immunopharmacol 8:1103-1111.
 Schreurs MW, Eggert AA, de Boer AJ, Figdor CG, Adema GJ (1999) Generation and functional characterization of mouse monocyte-derived dendritic cells. Eur J Immunol 29:2835-2841.
 Bowers WE, Berkowitz MR (1986) Differentiation of dendritic cells in cultures of rat bone marrow cells. J Exp Med 163:872-883.
 Inaba K, Inaba M., Romani N., Aya H., Deguchi M., Ikehara S., Muramatsu S., Steinman RM (1992) Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med 176:1693-1702.
 Talmor M, Mirza A, Turley S, Mellman I, Hoffman LA, Steinman RM (1998) Generation or large numbers of immature and mature dendritic cells from rat bone marrow cultures. Eur J Immunol 28:811-817.
 Bai L, Feuerer M, Beckhove P, Umansky V, Schirrmacher V (2002) Generation of dendritic cells from human bone marrow mononuclear cells: advantages for clinical application in comparison to peripheral blood monocyte derived cells. Int J Oncol 20:247-253.
 Lutz MB, Kukutsch N, Ogilvie AL, Rössner S, Koch F, Romani N, Schuler G (1999) An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods 223:77-92.