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Isolation and Characterization of Amniotic Fluid-Derived Extracellular Vesicles for Biomarker Discovery

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1885))

Abstract

Amniotic fluid, the fetal-protective liquid that fills the amniotic sac, represents a rich source of biomarkers. The diagnostic utility of amniotic fluid relies on the highly abundant maternal and fetal nucleic acid and proteomic content, which allows for the simultaneous determination of mother and fetal health status. Extracellular vesicles (ECVs) that are released by all cells and found in amniotic fluid could be harnessed to provide substantial clinically actionable data. ECVs are mediators of critical biological functions and reflect the health of the parent cell. Thus, ECVs released from cells in distress may provide important diagnostic information. Here, we describe a straightforward and optimized method for isolating ECVs from amniotic fluid. In addition, we validate our procedure through western blotting using antibodies targeting canonical ECV protein markers and via direct visualization using transmission electron microscopy.

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References

  1. Palmas F, Fattuoni C, Noto A et al (2016) The choice of amniotic fluid in metabolomics for the monitoring of fetus health. Expert Rev Mol Diagn 16(4):473–486. https://doi.org/10.1586/14737159.2016.1139456

    Article  CAS  PubMed  Google Scholar 

  2. Kolialexi A, Tounta G, Mavrou A et al (2011) Proteomic analysis of amniotic fluid for the diagnosis of fetal aneuploidies. Expert Rev Proteomics 8(2):175–185. https://doi.org/10.1586/epr.10.112

    Article  CAS  PubMed  Google Scholar 

  3. Evans MI, Andriole S, Evans SM (2015) Genetics: update on prenatal screening and diagnosis. Obstet Gynecol Clin N Am 42(2):193–208. https://doi.org/10.1016/j.ogc.2015.01.011

    Article  Google Scholar 

  4. Kamath-Rayne BD, Smith HC, Muglia LJ et al (2014) Amniotic fluid: the use of high-dimensional biology to understand fetal well-being. Reprod Sci 21(1):6–19. https://doi.org/10.1177/1933719113485292

    Article  PubMed  PubMed Central  Google Scholar 

  5. Urbanelli L, Buratta S, Sagini K et al (2015) Exosome-based strategies for Diagnosis and Therapy. Recent Pat CNS Drug Discov 10(1):10–27

    Article  CAS  Google Scholar 

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Correspondence to Alex J. Rai .

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Ebert, B., Rai, A.J. (2019). Isolation and Characterization of Amniotic Fluid-Derived Extracellular Vesicles for Biomarker Discovery. In: Levy, B. (eds) Prenatal Diagnosis. Methods in Molecular Biology, vol 1885. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8889-1_19

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  • DOI: https://doi.org/10.1007/978-1-4939-8889-1_19

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8887-7

  • Online ISBN: 978-1-4939-8889-1

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