不同细胞培养基对PAM212细胞产生TSLP的影响

To provide suitable cell culture media for the study of thymic stromal lymphopoietin (TSLP) production in PAM212 cells by comparing TSLP levels in four different cell culture media,TSLP levels in culture supernatant and cell viability were determined by ELISA and MTT assay.TSLP level in media of Dulbecco's modified Eagle's medium (DMEM) or Eagle's minimum essential medium (EMEM) was 5~6 times higher than that in Minimum essential medium alpha medium (MEMα) or RPMI1640,but the increase of TSLP expression in media of MEMα or RPMI1640 was higher than that in DMEM or EMEM.In order to investigate the effect of medium change on TSLP production,cell culture media were changed firstly,and then the cells were seeded,cultured for 24 h,and stimulated by 02F04 for 24 h.In order to compare TSLP production induced by the same stimuli in different cell culture media,PAM212 cells were seeded directly,cultured for 24 h,and stimulated by 02F04 at 10 μmol/L,TNF-α at 30 ng/mL or 12-O-tetradecanoylphorbol 13-acetate (TPA) at 3 nmol/L for 24 h,respectively.The results showed that TSLP production in PAM212 cells was directly related to the medium which was used from seeding,but not to that used in cell resuscitation or before seeding.TSLP levels induced by the three inducers were higher in the medium of DMEM,but the increase in TSLP expression was higher in MEMα.Besides,when PAM212 cells were cultured with fetal bovine serum (FBS)-free DMEM 10-h before or from the treatment of 02F04,TSLP production was significantly decreased,and the longer time FBS was removed for,the lower cell viability PAM212 cells had.Taken together,the levels of TSLP produced by PAM212 cells in various cell culture media were different.The medium of DMEM is suitable for a study in which a high level of TSLP in PAM212 cells without stimuli is required,whereas MEMα is suitable for the study on the mechanism of some stimuli on TSLP production in PAM212 cells because TSLP expression can be induced to a higher level.