[Study on rapid generation of dendritic cells from K562 cell line induced by A23187 alone].
OBJECTIVE
To explore a simple, rapid and efficient way to generate dendritic cells from leukemic cells.


METHODS
K562 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.


RESULTS
Microscopic 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).


CONCLUSIONS
A23187 treatment is a simple, rapid and efficient in vitro strategy for inducing dendritic cell from leukemic cells.
Zhao
Chun-ting
CT
Department of Hematology, Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, China.
Wang
Bao-zhong
BZ
Meng
Dong-mei
DM
Cao
Yong-xian
YX
Yang
Jie
J
Zhao
Xin-dong
XD
Chen
Bing
B
D000001:Calcimycin / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D020014:K562 Cells / Q000166:cytology / Q000187:drug effects*
D000001:Calcimycin