Isolation,induced culture and identification of immature dendritic cells in vitro
10.3969/j.issn.1000-484X.2017.07.018
- VernacularTitle:未成熟树突状细胞快速分离与体外诱导培养及其鉴定研究
- Author:
Na ZHU
;
Haitao WANG
;
Chang LI
;
Haixi LI
;
Yan GUO
;
Ningyi JIN
- Keywords:
Monocytes;
Immature dendritic cell;
Induced culture;
Appraisal
- From:
Chinese Journal of Immunology
2017;33(7):1043-1047
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a fast method for the generation of immature dendritic cells(DCs) from human peripheral blood mononuclear cells(hPBMCs) in vitro.Methods: High purity human CD14+ monocytes were collected using density Ficoll gradient centrifugation and MACS beads sorting system.iDCs(Immature DC) were induced after cultured with rhGM-CSF and rhIL-4 on the fourth day.Fluorescence activated cell sorting(FACS) was used to identify cell surface markers(CCR5) and capabilities of antigen uptake of iDCs on the fourth day.Ordinary optical microscope,scanning electron microscopy and transmission electron microscopy were used to observe surface and internal structure of iDCs on the fourth day of culture conditions.Results: FACS result shows that the purity of CD14+ monocytes collected from hPBMCs were more than 94%.The antigen uptake capability and CD195 of iDCs was detected on the fourth day of cultured conditions.Typical surface and internal structure characteristics of iDCs were observed.Conclusion: Rapid induction culture is an effective method for obtaining a large number of iDC with typical characteristics in vitro,and can be used for further experimental study.
