Proliferation of Dendritic Cells from Human Cord Blood in Vitro

Objective To investigate the proliferation and functions of dendritic cells derived from human cord blood progenitor cells in vitro.Methods The mononuclear cells were prepared from cord blood with Ficoll-hypaque centrifugation method,then induced with the recombinant cytokines hGM-CSF(50 ng/ml),hIL-4(10 ng/ml) and hTNF-α(50 ng/ml) for two weeks.We observed the DCs growth under the microscope and determined DCs phenotypes by flow cytometry.The capacity of DCs to activate T cell-depent anti-tumor immune responses was tested by MTT.Results The dendritic cells cultured in vitro turned into suspensive growth from adhesive situation at 6 d.Then the number of DCs increased continuously and the cells showed the irregular morphologic appearance of DCs with veiled edges on the 12th day.It was showed by flowcytometry that the mature DCs expressed the relatively specific marker such as HLA-DR,CD80,CD83,CD1a,CD11c and CD123.T cells activated by DCs showed magnificent cytotoxicity on two different kinds of tumor cells.Conclusion The dendritic cells can be induced from human cord blood through the cultivation with rhGM-CSF,rhIL-4 and rhTNF-α in vitro.