NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis
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Postmortem RNA and protein stability in perinatal human lungs.

Author(s): De Paepe ME, Mao Q, Huang C, Zhu D, Jackson CL, Hansen K

Publication: Diagn Mol Pathol, 2002, Vol. 11, Page 170

PubMed ID: 12218457 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to evaluate the suitability of RNA and protein extracted from postmortem perinatal lung tissue for molecular analyses. The authors compared tissue from stillbirths and live-births that had been snap frozen or preserved with RNAlater, and after variable postmortem intervals (PMIs).

Conclusion of Paper

RNA and protein yield and integrity were greater in specimens preserved with RNAlater compared to liquid nitrogen, and was not affected by PMI (up to 93 h), although live versus still birth and maceration influenced RNA integrity and mRNA and protein expression. The authors conclude that RNA and protein extracted from postmortem lung from live and non-macerated stillborn infants can successfully be used in molecular analysis.

Studies

  1. Study Purpose

    The purpose of this study was to assess the effects of stillbirth and postmortem interval on the quality of RNA and protein extracted from perinatal (median gestational age 23 weeks) lung tissue that had been preserved via snap freezing or RNAlater. Postmortem interval for stillbirths was calculated from delivery and did not include postmortem time in utero.

    Summary of Findings:

    RNA yield was significantly greater in specimens preserved in RNAlater and stored at -20 degrees C compared to those snap frozen and stored at -80 degrees C regardless of live versus still birth or PMI (up to 93h). RNA integrity was comparable among specimens preserved with RNAlater or liquid nitrogen, although specimens collected from live births were significantly greater than those from still births. Maceration further affected RNA quality, while PMI and length of life did not. Beta-actin and surfactant protein-B (SP-B) mRNA levels were comparable among preservation methods, although were significantly lower in macerated stillborns. Protein yield was greater from specimens preserved in RNAlater than snap-frozen in liquid nitrogen and was greater from live-born than stillborn lungs for RNAlater-preserved tissue. Postmortem interval did not influence protein yield. No statistical differences were observed in Western blot evaluation of protein immunoreactivity regardless of preservation method, live-born or stillborn specimens, or postmortem interval.

    Biospecimens
    Preservative Types
    • RNAlater
    • Frozen
    Diagnoses:
    • Autopsy
    • Not specified
    Platform:
    AnalyteTechnology Platform
    RNA Spectrophotometry
    Protein Western blot
    Protein Lowry protein assay
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Preaquisition Postmortem interval 0 - 4 h
    5 - 9 h
    10 - 14 h
    15 - 19 h
    20 - 24 h
    25 - 41 h
    Biospecimen Preservation Type of fixation/preservation RNAlater
    Snap frozen
    Preaquisition Diagnosis/ patient condition Live-born
    Stillborn, non-macerated
    Stillborn, macerated
    Storage Storage temperature -20 degrees C
    -80 degrees C
    View more

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