Study on rapid generation of dendritic cells from K562 cell line induced by A23187 alone.
- Author:
Chun-ting ZHAO
1
;
Bao-zhong WANG
;
Dong-mei MENG
;
Yong-xian CAO
;
Jie YANG
;
Xin-dong ZHAO
;
Bing CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Calcimycin; pharmacology; Cell Differentiation; drug effects; Cells, Cultured; Coculture Techniques; Dendritic Cells; cytology; immunology; metabolism; Humans; K562 Cells; cytology; drug effects
- From: Chinese Journal of Hematology 2005;26(7):408-412
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore a simple, rapid and efficient way to generate dendritic cells from leukemic cells.
METHODSK562 cells were cultured with calcium ionosphere A23187 alone, A23187 plus GM-CSF, or a DC differentiation cocktail consisting of GM-CSF, IL-4 and TNF-alpha, respectively. The expression of surface markers of induced DCs was analyzed by flow cytometry. The K562-DCs stimulating the proliferation of allo-genetic naive T cells and inducing cytotoxicity of T cells were determined by MTT assay.
RESULTSMicroscopic examination revealed that under all the three culture conditions, K562 cells became displaying DC morphology. At 72 hours in the two culture systems containing A23187, there were higher proportions of cells with dendritic morphology [(69.5 +/- 17.2)% and (73.1 +/- 13.9)%, respectively] than that in the cocktail system [(28.5 +/- 12.3)%] (P < 0.05). And the same did when cultured for 7 days [(69.5 +/- 17.2)%, (73.1 +/- 13.9)% respectively vs (51.2 +/- 10.7)%, P < 0.05]. In the 7-day cultures, the percentage of CD1a expressing cells was lower [(8.2 +/- 2.3)% and (10.3 +/- 5.1)% vs (17.2 +/- 1.6)%, respectively] while the CD83 expressing cells was higher [(85.6 +/- 8.8)% and (82.4 +/- 9.1)% vs (77.4 +/- 12.9)%, respectively] compared with that in the cocktail system (P < 0.05). No significant difference was found in the allogeneic T cell proliferation response and induced T cell cytotoxicity between A23187 containing and cocktail groups (P > 0.05).
CONCLUSIONSA23187 treatment is a simple, rapid and efficient in vitro strategy for inducing dendritic cell from leukemic cells.