optimized method for ex vivo generation of dendritic cells

ABSTRACT

Dendritic cells are the most potent antigen presenting cells. They capture, process and present antigens to T cells and secrete various cytokines and soluble factors to initiate adaptive immune responses. Dendritic cells are also important in induction of immunological tolerance to self- antigens. Due to their crucial role in immune responses during infections, cancers, transplantations, allergies, and autoimmune diseases, they have become important targets for many biological and clinical studies. Usually large numbers of dendritic cells is essential for these studies; however, small numbers of these cells in blood and tissues makes the isolation very difficult. Therefore, In vitro generation of these cells is useful for research and clinical applications. The aim of this study was in vitro generation of dendritic cells from bone marrow-hematopoietic progenitor cells using a simple and efficient method. Murine bone marrow cells were cultured in medium supplemented with Granulocyte-macrophage colony-stimulating factor and Interleukin 4 without depletion of any cell population for 9 days. Fresh medium containing cytokines was added every 3 days. Analysis of the morphology and immunophenotype of cultured cells showed the generation of dendritic cells from day 2 of the culture period. But, the number of cells that possess morphological characteristics and typical cell surface markers [CD11c, MHC-II and CD86] of dendritic cells was elevated by increasing the culture period. The purity of dendritic cells was 86% by the end of 9 days culture. This method can be used as an efficient method for ex vivo generation of dendritic cells