GENERATION AND MATURATION OF BONE MARROW-DERIVED DENDRITIC CELLS. A COMPARATIVE STUDY BETWEEN FRESH AND CRYOPRESERVED CELLS

Abstract

Dendritic cells (DCs) are essential for the induction of the adaptive immune responses, by activating naive T-cells and orchestrating antigen-specific T cell differentiation based on signals received from pathogens. Therefore in vaccine development, dendritic cell based in vitro assays could allow the investigation of many vaccine candidates and selection of the best antigens and adjuvants during the process of development. However, to establish a highly efficient and reproducible protocol and to reduce the number of animals used for experiments, the use of cryopreserved precursors is desirable. Here we compared the differentiation of freshly harvested and cryopreserved mouse bone marrow cells (BM) cultured under the same condition and we found that cryopreserved BM can effectively be differentiated to bone marrow-derived dendritic cells (BMDCs) in the presence of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF). High purity of immature BMDCs was obtained in both conditions, even though cryopreservation precursors showed a slowdown in the initial development and a lower yield, up to 10 times than from fresh cultures. Furthermore, BMDCs obtained from both fresh or cryopreserved cells undergo maturation in response to lipopolysaccharides (LPS), triggering a high-level expression of costimulatory molecules such as CD86 and MHC-II.