Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM...Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM-CSF, TNF-a, IL-1b, 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 wegenerated from frozen PBMCs were able to prime CD8 T cells into long term IFN-g producing peptide specific CTL. Conclusion: The DCs we developed from cryopreservedPBMC were fully mature and had the capability tostimulate immune reaction. Thus, we developed a method to generate functional mature DC from cryopreservedPBMC.展开更多
文摘Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM-CSF, TNF-a, IL-1b, 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 wegenerated from frozen PBMCs were able to prime CD8 T cells into long term IFN-g producing peptide specific CTL. Conclusion: The DCs we developed from cryopreservedPBMC were fully mature and had the capability tostimulate immune reaction. Thus, we developed a method to generate functional mature DC from cryopreservedPBMC.
