In vitro culture and expansion of dendritic cells derived from human umbilical cord blood

Yang Bo, Qi Yanchao, Lu Minying

1. Department of Bioengineering, Guangzhou Cancer Hospital 2. Department of Bioengineering, Guangzhou Cancer Hospital 3. Department of Bioengineering, Guangzhou Cancer Hospital 5100954.5100955.510095

Abstract: Objective To investigate the function of dendritic cells (DCs) derived from human umbilical cord blood in vitro. Methods The interface cells were obtained by centrifugation with Ficoll Hypaque and cultured for 2h to obtain mononuclear cells, which were induced by recombinant hGM CSF (50 ng/ml) hIL 4( 10ng/ml) hTNFα (50 ng/ml) for 14 days in vitro. During the development of DCs, the growth of DCs was observed under light microscope, and the phenotype of DCs was detected by flow cytometry. MTT assay was used to detect the killing of T cells activated by DCs to tumor cells. Results On the 6th day, DCs cultured in vitro changed from adherent state to suspended burr-like cells. With the extension of culture time, the number of such cells increased. On the 12th day, DCs were irregular burr-like and typical dendritic cell morphology. Flow cytometry showed that mature DCs expressed HLA DR, CD 80, CD83, CD 1a, CD 11c, CD12 3 at high levels. Activated T cells produced significant damage to both tumor cells derived from different tissues. Conclusion DCs can be induced by human umbilical cord blood cultured in vitro with rhGM CSF rhIL 4 hTNF α.

Key words:Dendritic cells;Cord blood;Recombinant human granulocyte-macrophage colony stimulating factor;Recombinant human interleukin-4;Recombinant human tumor necrosis factor alpha;

• Album: Medical and Health Technology
• Special Topic: Oncology
• Classification number: R73-3

In vitro culture and expansion of dendritic cells from human umbilical cord blood