2-mercaptoethanol serum supplement

Growth medium RPMI-1640 with L-glutamine (Irvine Scientific Co., Irvine, CA), supplemented with 10% heat-inactivated fetal bovine serum, lOIU/mL penicillin, lOOpg/mL streptomycin (PEST), 50pM2-mercaptoethanol, 292 igl mL L-glutamine, and lOmM HEPES buffer (all from Sigma-Aldrich). 

Low serum-containing medium Iscove s modified Dulbecco s medium (IMDM) (Sigma Chemical Company, St Louis, MO, USA) supplemented with 2% foetal bovine serum and 5 x 1(L M p-mercaptoethanol... 

RIO cell culture medium (RPMI 1640, supplemented with 10% fetal calf serum (FCS), 2 mM 1-glutamin, 100 U/mL peniciUin, 100 pg/mL streptomycin and 50 pM 2-mercaptoethanol (all Invitrogen)). 

Fig. 1. Effect of 3-methoxybenzamide and cyclosporin A on thymidine incorporation by thymus organ cultures. Thymic lobes were removed from 14-day-old mouse embryos and cultured on the surface of a nucleopore filter on gelatin foam squares in Dulbecco s Modified Eagles s Medium supplemented with nonessential amino acids (Flow), 50 iM 2-mercaptoethanol and 10% foetal calf serum. After 5 days at 37°C, the lobes were transferred to hanging-drop cultures in Terasaki plates and H]-thymidine was added to a final concentration of 2 juCi ml. After a further 15 h at 37°C the thymic lobes were washed, disaggregated with trypsin-EDTA and harvested for determination of the level of incorporation of [ H]-thymidine. In drug-treated cultures, either 1 jug nil cyclosporin A or 2.5 mM 3-methoxybenzamide was included in the medium (both before and during the [ H]-thymidine pulse). Means and SE of three experiments, each of which involved at least four thymuses, are shown...

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