In vitro cultivation of dendritic cells with serum-free medium / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 985-989, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-282748
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Dendritic Cells
/
Recombinant Proteins
/
Monocytes
/
T-Lymphocytes
/
T-Lymphocytes, Cytotoxic
/
Granulocyte-Macrophage Colony-Stimulating Factor
/
Interleukin-4
/
Culture Media, Serum-Free
/
Cell Culture Techniques
Type of study:
Practice guideline
Limits:
Humans
Language:
Chinese
Journal:
Journal of Experimental Hematology
Year:
2006
Type:
Article
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