In vitro cultivation of dendritic cells with serum-free medium.
- Author:
Huai-Dong ZHANG
1
;
Zhen-Lan SONG
;
Wei-Ping LI
Author Information
1. Department of Hematology, Shanton Central Hospital, Shanton 310003, China. zhd1977@163.com
- Publication Type:Journal Article
- MeSH:
Cell Culture Techniques;
Culture Media, Serum-Free;
Dendritic Cells;
cytology;
Granulocyte-Macrophage Colony-Stimulating Factor;
pharmacology;
Humans;
Interleukin-4;
pharmacology;
Monocytes;
cytology;
Recombinant Proteins;
T-Lymphocytes;
cytology;
T-Lymphocytes, Cytotoxic
- From:
Journal of Experimental Hematology
2006;14(5):985-989
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the protocol in vitro to incubate the dendritic cell (DC) derived from peripheral blood monocytes using serum-free medium X-VIVO 20. Peripheral blood monocytes from healthy donors were treated with 100 ng/ml GM-CSF and 500 U/ml IL-4, respectively. After cultivation for 6 days, they were treated with 100 ng/ml calcium ionophore A23187. After cultivation for 24 hours the cellular morphology was observed under invert microscope, the surface markers were analyzed by flow cytometry, the proliferation of allogenetic T cells was detected by MTT colorimetry, the specific cytotoxicity of T cells primed with DC was examined by MTT assay. The results showed that in all three groups with serum-free, fetal calf serum (FCS) and human AB serum mediums, cells displayed characteristic morphological features of DC. Simultaneously CD14 expression was decreased, and CD83, HLA-DR and CDw123 expression were increased on these cells. In addition, DCs cultured with these methods could evidently stimulate the proliferation of allogenetic T cell. As compared with the two controls of serum containing groups, the cultured cells in the serum-free groups showed almost the same allo-stimulatory capability and cellular morphology and surface markers, and T lymphocytes primed with the culture-derived DC exhibited the similar killing activity to K562 (P > 0.05). It is concluded that there is no significance in DC numbers, morphology, epitope and ability to stimulate the proliferation of allogenetic T cells between DC induced by serum-free X-VIVO 20 medium and DC induced by serum-contained medium. DC cultured and induced by serum-free medium is worth using in practice widely.
