Optimization of cytokine stability in stored amniotic fluid

Amy E. Porter, Jonathan Auth, Michele Prince, Alessandro Ghidini, Douglas E. Brenneman, Catherine Y. Spong

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

OBJECTIVE: Many studies use stored amniotic fluid samples to assay cytokines and other proteins for outcome-based research; however, there is little information on the optimal methods of storage. The objective of our study was to evaluate cytokine stability in amniotic fluid stored at different temperatures both with and without a proteolytic enzyme inhibitor. STUDY DESIGN: Patients undergoing midtrimester genetic amniocentesis for routine indications gave consent for the study. After the sample was centrifuged, the acellular portion of the sample was mixed to homogeneity and aliquoted in 0.5-mL increments and stored for 1 year at 4°C, -20°C, and -80°C with and without the protease inhibitor aprotinin. Enzyme-linked immunoassays for angiogenin, interleukin-6, and vascular endothelial growth factor were performed simultaneously on each aliquot. RESULTS: Thirty samples were assayed for each storage condition. Results were calculated as the percentage of its own sister aliquot stored at -80°C without aprotinin. In all samples, there was a significant relation between storage temperature and cytokine levels, with the lowest levels found at 4°C and the highest at -80°C (angiogenin, P = .004; interleukin-6, P < .001; vascular endothelial growth factor, P = .02). The addition of aprotinin improved stability only for angiogenin at all temperatures (all P < .05). CONCLUSIONS: Degradation of cytokines occurs when amniotic fluid samples are stored for prolonged periods at temperatures greater than -80°C. The addition of a protease inhibitor helps stem the degradation of some cytokines.

Original languageEnglish (US)
Pages (from-to)459-462
Number of pages4
JournalAmerican journal of obstetrics and gynecology
Volume185
Issue number2
DOIs
StatePublished - Jan 1 2001
Externally publishedYes

Fingerprint

Amniotic Fluid
Aprotinin
Cytokines
Temperature
Protease Inhibitors
Vascular Endothelial Growth Factor A
Interleukin-6
Amniocentesis
Second Pregnancy Trimester
Enzyme Inhibitors
Immunoenzyme Techniques
Siblings
Peptide Hydrolases
Outcome Assessment (Health Care)
angiogenin
Proteins

Keywords

  • Amniotic fluid
  • Angiogenin
  • Cytokines
  • Interleukin-6
  • Stability
  • Vascular endothelial growth factor

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Optimization of cytokine stability in stored amniotic fluid. / Porter, Amy E.; Auth, Jonathan; Prince, Michele; Ghidini, Alessandro; Brenneman, Douglas E.; Spong, Catherine Y.

In: American journal of obstetrics and gynecology, Vol. 185, No. 2, 01.01.2001, p. 459-462.

Research output: Contribution to journalArticle

Porter, Amy E. ; Auth, Jonathan ; Prince, Michele ; Ghidini, Alessandro ; Brenneman, Douglas E. ; Spong, Catherine Y. / Optimization of cytokine stability in stored amniotic fluid. In: American journal of obstetrics and gynecology. 2001 ; Vol. 185, No. 2. pp. 459-462.
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AB - OBJECTIVE: Many studies use stored amniotic fluid samples to assay cytokines and other proteins for outcome-based research; however, there is little information on the optimal methods of storage. The objective of our study was to evaluate cytokine stability in amniotic fluid stored at different temperatures both with and without a proteolytic enzyme inhibitor. STUDY DESIGN: Patients undergoing midtrimester genetic amniocentesis for routine indications gave consent for the study. After the sample was centrifuged, the acellular portion of the sample was mixed to homogeneity and aliquoted in 0.5-mL increments and stored for 1 year at 4°C, -20°C, and -80°C with and without the protease inhibitor aprotinin. Enzyme-linked immunoassays for angiogenin, interleukin-6, and vascular endothelial growth factor were performed simultaneously on each aliquot. RESULTS: Thirty samples were assayed for each storage condition. Results were calculated as the percentage of its own sister aliquot stored at -80°C without aprotinin. In all samples, there was a significant relation between storage temperature and cytokine levels, with the lowest levels found at 4°C and the highest at -80°C (angiogenin, P = .004; interleukin-6, P < .001; vascular endothelial growth factor, P = .02). The addition of aprotinin improved stability only for angiogenin at all temperatures (all P < .05). CONCLUSIONS: Degradation of cytokines occurs when amniotic fluid samples are stored for prolonged periods at temperatures greater than -80°C. The addition of a protease inhibitor helps stem the degradation of some cytokines.

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