Open Access

ARTICLE

Stability of cytokines in supernatants of stimulated mouse immune cells

Ozbey G¹, Gorczynski R², Erin N¹

1 Department of Medical Pharmacology, School of Medicine, Akdeniz University, Dumlupınar Street, Kampus, 07070, Antalya, Turkey
2 The Toronto Hospital, University Health Network, Toronto, Canada; 2-805, MaRS Tower, 101 College Street, University Health Network and TheToronto Hospital, Toronto, Ontario, Canada M5G1L7

* Corresponding Authors: ; ;

European Cytokine Network 2014, 25(2), 30-34. https://doi.org/10.1684/ecn.2014.0353

Abstract

Measurements of cytokines in cell culture supernatants are widely used to evaluate the immune response. Cytokine levels in secretomes are usually quantified using enzyme-linked immunosorbent assays (ELISA), which have easy, sensitive, specific, rapid, cost-effective, and reproducible protocols. To our knowledge, the stability of cytokines in secretomes has not been hitherto investigated. We present data that involve; time-dependent changes during storage at +4℃, and the effects of freeze-thaw cycles in samples frozen at -80℃, instant freezing of samples with liquid nitrogen, and addition of protease inhibitors on the stability of certain cytokines (TNF-α, MIP-2, IFN-γ, IL-6, IL-10, IL-17A), in secrotomes of spleen and lymph nodes from tumor-bearing animals. Our results show that IL-6 remains stable, MIP-2, IFN-γ and IL-10 are somewhat stable, while TNF-α and IL-17A are degradable cytokines: instant freezing by liquid nitrogen or adding protease inhibitor does not preserve the stability of these cytokines. From these results it can be concluded that, if possible, TNF-α measurements should be perform in fresh samples, and IL-17A and IL-10 samples can be stored at -80℃, but should be used at the first thaw.

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Keywords

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