*2.11. Detection of Cytokines in Cell Culture Fluids by Sandwich ELISA*

Measurement of cytokine levels (IFN-γ, TNF-α, IL-2, IL-6, IL-10, IL-12) was performed by ELISA in conditioned medium from selection of the MSC cells transfected with pcNS3-NS5B. The concentration of IFN-γ was also determined in medium from the splenocytes stimulated for four days. We used the Mouse IL-6 ELISA development kit (HRP), Mouse IFN-γ ELISA development kit (HRP), Mouse TNF-α ELISA development kit (HRP) (Mabtech, Stockholm, Sweden), Mouse IL-10 DuoSet ELISA, and Mouse IL-12 p70 Duoset ELISA (R&D Systems, Minneapolis, MN, USA). The detection sensitivity for IL-6 was 10 pg/mL, for IFN-γ and TNF-α—2 pg/mL, for IL-2—4 pg/mL, for IL-10 and IL-12—30 pg/mL. The concentrations of cytokines were determined from the calibration curves of standard samples.
