Culture and functional identification of macrophages from human peripheral blood
10.3969/j.issn.1000-484X.2015.01.018
- VernacularTitle:人外周血巨噬细胞培养及功能鉴定
- Author:
Weiming LIN
;
Ailing DAI
;
Huifang YIN
;
Xiaoyan YANG
- Publication Type:Journal Article
- Keywords:
Macrophages;
Magnetic activated cell sorting;
Culture;
Functional identification
- From:
Chinese Journal of Immunology
2015;(1):86-89
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To isolate monocytes from human peripheral blood mononuclear cells( PBMC) ,induce macrophages,and identify the function of macrophages.Methods:Monocytes were isolated from PBMC using magnetic activated cell sorting( MACS) anti-CD14 microbead.Sorted CD14+and CD14-cells were checked by flow cytometer to evaluate the efficiency of sorting.The sorted CD14+cells were cultured in IMDM media with 10%human AB serum and 10 ng/ml M-CSF for 7 days to generate macrophages,which were identified by morphological features and phagocytosis function.Results:A high purity of monocytes was obtained by MACS anti-CD14 microbead.The percentage of CD14+cells was 10% and 85.8% before and after sorting, respectively.The macrophages were approximately 40-45 μm in maximum diameter and had the fried egg colony morphological features after 7 days culture.The lymphoma ( Raji) cells were efficiently engulfed by macrophages.Conclusion: The high purity of CD14+monocytes is isolated from PBMC and monocyte-derived macrophages efficiently engulfed lymphoma cells.
