Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) and producing DCs from cryopreserved PBMCs. Methods: Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM-CSF, TNF-α, IL-1β, IL-6, pgE2 and LPS. The phenotype of the resultant DCs was investigated by flow cytometry. The functions of the resultant DCs were verified by Elispot assay. Results: The resultant DCs expressed high levels of HLA ABC, HLA DR, costimulatory molecules and the DC maturation marker CD83. The mature DCs we generated from frozen PBMCs were able to prime CD8 T cells into long term IFN-γ producing peptide specific CTL. Conclusion: The DCs we developed from cryopreserved PBMC were fully mature and had the capability to stimulate immune reaction. Thus, we developed a method to generate functional mature DC from cryopreserved PBMC.
- Cytotoxic T lymphocyte (CTL)
- Dendritic cell (DC)
- Peripheral blood mononuclear cell (PBMC)