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Delay in processing porcine whole blood affects cytokine expression
J C Duvigneau1, R T Hartl, M Teinfalt
1Institute of Medical Chemistry, Veterinary University, Veterinärplatz 1, A-1210 Vienna, Austria. catharina.duvigneau@vu-wien.ac.at
Journal of Immunological Methods
|December 31, 2002
Summary
Porcine blood sample storage affects cytokine messenger ribonucleic acid (mRNA) levels. Processing within 2 hours is recommended, but temperature control can mitigate changes for specific cytokines.
Area of Science:
- Veterinary Immunology
- Molecular Biology
- Biochemistry
Background:
- Cytokine messenger ribonucleic acid (mRNA) quantification is crucial for studying immune cell activation.
- Sample collection and processing methods significantly impact cytokine mRNA levels.
- Standardized and timely processing of blood samples is generally advised.
Purpose of the Study:
- To investigate the impact of storage temperature (room temperature and 4°C) and duration (1-24 hours) on cytokine mRNA levels in porcine whole blood.
- To determine optimal processing times and conditions to ensure accurate cytokine expression analysis.
Main Methods:
- Porcine whole blood samples (n=8) were stored under different conditions.
- Cytokine mRNA levels (IL-1α, IL-2, IL-6, IL-8, IL-10, IFN-γ, IL-2R) were quantified using simultaneous reverse transcription PCR (RT-PCR).
- Expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a reference housekeeping gene.
Main Results:
- Storage beyond 1 hour led to significant changes in individual cytokine mRNA levels.
- Monokine expression (IL-1α, IL-6, IL-10) increased at both room temperature and 4°C.
- IL-8 mRNA increased at room temperature, while IFN-γ mRNA increased at 4°C.
Conclusions:
- Porcine blood samples require processing within 2 hours to avoid artificial stimulation of cytokine expression.
- Storage temperature can selectively influence specific cytokine mRNA levels, offering potential mitigation strategies for certain analyses.