Induction and culturing of dendritic cells derived from human peripheral blood monocytes in vitro
- VernacularTitle:人类外周血来源树突状细胞的体外诱导培养
- Author:
Jianwu LIU
;
Bing WANG
;
Jiwen SONG
;
Huiqing CHEN
- Publication Type:Journal Article
- Keywords:
Dendritic cells;
Culture;
Phenotype
- From:
Cancer Research and Clinic
2007;19(z1):17-18,22
- CountryChina
- Language:Chinese
-
Abstract:
Objective To generate high purity and maturity DC from human peripheral blood monocytes in vitro.Methods PBMC were isolated directly from whole blood by density gradient centrifugation and purified by collecting the attached cell,DC were then generated by induction and culturing PBMC for five days with RPMI1640 medium containing rhGM-CSF and rhIL-4 in vitro,and under the condition of 37 ℃,5% CO2.On the fifth day,rhTNF-α was added into DCs cultures,which were then incubated for three additional days.The morphology was monitored by light microscopy,and the phenotypes were determined by FCM.Results After eight days of culture,the cells developed typical and significant dendritic morphology and plenty of cells expressed CD1a, CD80 and CD83,features of DC.Including(78.07±9.43)%CD1a,(60.11±20.50)% CD80 and(46.82±14.15)% CD83 were expressed.About (3.12±1.30)x106 DC cells were derived from 40ml human peripheral blood.Conclusion The way to generate DCs is simple and easy.The DCs produced by this method acquired high purity and maturity antigenic characteristics of DCs.
