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Chapter 19 Cryopreservation Effect on Genetic Function: Neonatal Outcomes.

Vanesa Robles1, Marta F Riesco2, David G Valcarce2

  • 1Department of Molecular Biology, Cell Biology Area, University of León, Campus de Vegazana s/n, León, 24071, León, Spain. robles.vanesa@gmail.com.

Methods in Molecular Biology (Clifton, N.J.)
|April 20, 2017
PubMed
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Cryopreservation can alter gametes and embryos at the molecular level, potentially impacting DNA and epigenetics. New diagnostic tools are needed to refine cryopreservation methods and prevent undesirable changes.

Area of Science:

  • Reproductive biology
  • Molecular biology
  • Cryobiology

Background:

  • Cryopreservation is essential for clinical applications like assisted reproduction.
  • Established cryopreservation techniques can induce molecular alterations in gametes and embryos.
  • These alterations include changes in messenger RNA, epigenome, and DNA damage.

Purpose of the Study:

  • To describe a protocol for quantifying cryopreservation-induced lesions.
  • To aid in the development of optimized cryopreservation protocols.
  • To minimize undesirable molecular changes in reproductive cells.

Main Methods:

  • Quantification of DNA lesions.
  • Utilizing a previously published protocol by Rothfuss.
  • Application to cryopreserved gametes and embryos.
Keywords:
CryopreservationDNA damageDNA extractionDNA lesionsQuantitative polymerase chain reaction

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Main Results:

  • The protocol allows for the measurement of specific molecular damage induced by cryopreservation.
  • Identifies the extent of DNA lesions resulting from the cryopreservation process.

Conclusions:

  • Accurate quantification of cryopreservation-induced damage is crucial.
  • This protocol provides a means to assess and potentially mitigate molecular alterations.
  • Further research into diagnostic tools can improve assisted reproductive technologies.