Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification

Bence Somoskoi, Nicola A. Martino, Rosa A. Cardone, Giovanni M. Lacalandra, Maria E. Dell'Aquila, S. Cseh

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: The ability to cryopreserve mammalian embryos has become an integral part of assisted reproduction, both in human and veterinary medicine. Despite differences in the size and physiological characteristics of embryos from various species, the embryos have been frozen by either of two procedures: slow freezing or vitrification. The aim of our study was to compare the effect of slow freezing and vitrification to the chromatin structure, energy status and reactive oxygen species production of mouse morulae and blastocysts. Methods: Mouse morulae and blastocysts were randomly allocated into vitrification, slow freezing and control groups. For slow freezing, Dulbecco phosphate buffered saline based 10% glicerol solution was used. For vitrification, G-MOPS™ based solution supplemented with 16% ethylene glycol, 16% propylene glycol, Ficoll (10mg/ml) and sucrose (0.65mol/l) was used. After warming, the chromatin integrity, mitochondrial distribution pattern and energy/oxidative status were compared among groups. Results: Cryopreservation affected chromatin integrity at a greater extent at the morula than the blastocyst stage. Chromatin damage induced by slow freezing was more relevant compared to vitrification. Slow freezing and vitrification similarly affected mitochondrial distribution pattern. Greater damage was observed at the morula stage and it was associated with embryo grade. Cryopreservation altered the quantitative bioenergy/redox parameters at a greater extent in the morulae than in the blastocysts. Effects induced by slow freezing were not related to embryo grade or mitochondrial pattern, as affected embryos were of all grades and with both mitochondrial patterns. However, effects induced by vitrification were related to mitochondrial pattern, as only embryos with homogeneous mitochondrial pattern in small aggregates had reduced energy status. Conclusions: This study shows for the first time the joint assessment of chromatin damage and mitochondrial energy/redox potential in fresh and frozen mouse embryos at the morula and blastocyst stage, allowing the comparison of the effects of the two most commonly used cryopreservation procedures.

Original languageEnglish
Article number22
JournalReproductive Biology and Endocrinology
Volume13
Issue number1
DOIs
Publication statusPublished - Mar 24 2015

Fingerprint

Vitrification
Morula
Blastocyst
Freezing
Chromatin
Oxidation-Reduction
Embryonic Structures
Cryopreservation
Mammalian Embryo
Ficoll
Propylene Glycol
Veterinary Medicine
Ethylene Glycol
Reproduction
Sucrose
Reactive Oxygen Species
Joints
Phosphates
Control Groups

Keywords

  • Mitochondria
  • Mouse embryos
  • Nuclear chromatin
  • Reactive oxygen species
  • Slow freezing
  • Vitrification

ASJC Scopus subject areas

  • Developmental Biology
  • Endocrinology
  • Reproductive Medicine

Cite this

Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification. / Somoskoi, Bence; Martino, Nicola A.; Cardone, Rosa A.; Lacalandra, Giovanni M.; Dell'Aquila, Maria E.; Cseh, S.

In: Reproductive Biology and Endocrinology, Vol. 13, No. 1, 22, 24.03.2015.

Research output: Contribution to journalArticle

Somoskoi, Bence ; Martino, Nicola A. ; Cardone, Rosa A. ; Lacalandra, Giovanni M. ; Dell'Aquila, Maria E. ; Cseh, S. / Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification. In: Reproductive Biology and Endocrinology. 2015 ; Vol. 13, No. 1.
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AU - Somoskoi, Bence

AU - Martino, Nicola A.

AU - Cardone, Rosa A.

AU - Lacalandra, Giovanni M.

AU - Dell'Aquila, Maria E.

AU - Cseh, S.

PY - 2015/3/24

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N2 - Background: The ability to cryopreserve mammalian embryos has become an integral part of assisted reproduction, both in human and veterinary medicine. Despite differences in the size and physiological characteristics of embryos from various species, the embryos have been frozen by either of two procedures: slow freezing or vitrification. The aim of our study was to compare the effect of slow freezing and vitrification to the chromatin structure, energy status and reactive oxygen species production of mouse morulae and blastocysts. Methods: Mouse morulae and blastocysts were randomly allocated into vitrification, slow freezing and control groups. For slow freezing, Dulbecco phosphate buffered saline based 10% glicerol solution was used. For vitrification, G-MOPS™ based solution supplemented with 16% ethylene glycol, 16% propylene glycol, Ficoll (10mg/ml) and sucrose (0.65mol/l) was used. After warming, the chromatin integrity, mitochondrial distribution pattern and energy/oxidative status were compared among groups. Results: Cryopreservation affected chromatin integrity at a greater extent at the morula than the blastocyst stage. Chromatin damage induced by slow freezing was more relevant compared to vitrification. Slow freezing and vitrification similarly affected mitochondrial distribution pattern. Greater damage was observed at the morula stage and it was associated with embryo grade. Cryopreservation altered the quantitative bioenergy/redox parameters at a greater extent in the morulae than in the blastocysts. Effects induced by slow freezing were not related to embryo grade or mitochondrial pattern, as affected embryos were of all grades and with both mitochondrial patterns. However, effects induced by vitrification were related to mitochondrial pattern, as only embryos with homogeneous mitochondrial pattern in small aggregates had reduced energy status. Conclusions: This study shows for the first time the joint assessment of chromatin damage and mitochondrial energy/redox potential in fresh and frozen mouse embryos at the morula and blastocyst stage, allowing the comparison of the effects of the two most commonly used cryopreservation procedures.

AB - Background: The ability to cryopreserve mammalian embryos has become an integral part of assisted reproduction, both in human and veterinary medicine. Despite differences in the size and physiological characteristics of embryos from various species, the embryos have been frozen by either of two procedures: slow freezing or vitrification. The aim of our study was to compare the effect of slow freezing and vitrification to the chromatin structure, energy status and reactive oxygen species production of mouse morulae and blastocysts. Methods: Mouse morulae and blastocysts were randomly allocated into vitrification, slow freezing and control groups. For slow freezing, Dulbecco phosphate buffered saline based 10% glicerol solution was used. For vitrification, G-MOPS™ based solution supplemented with 16% ethylene glycol, 16% propylene glycol, Ficoll (10mg/ml) and sucrose (0.65mol/l) was used. After warming, the chromatin integrity, mitochondrial distribution pattern and energy/oxidative status were compared among groups. Results: Cryopreservation affected chromatin integrity at a greater extent at the morula than the blastocyst stage. Chromatin damage induced by slow freezing was more relevant compared to vitrification. Slow freezing and vitrification similarly affected mitochondrial distribution pattern. Greater damage was observed at the morula stage and it was associated with embryo grade. Cryopreservation altered the quantitative bioenergy/redox parameters at a greater extent in the morulae than in the blastocysts. Effects induced by slow freezing were not related to embryo grade or mitochondrial pattern, as affected embryos were of all grades and with both mitochondrial patterns. However, effects induced by vitrification were related to mitochondrial pattern, as only embryos with homogeneous mitochondrial pattern in small aggregates had reduced energy status. Conclusions: This study shows for the first time the joint assessment of chromatin damage and mitochondrial energy/redox potential in fresh and frozen mouse embryos at the morula and blastocyst stage, allowing the comparison of the effects of the two most commonly used cryopreservation procedures.

KW - Mitochondria

KW - Mouse embryos

KW - Nuclear chromatin

KW - Reactive oxygen species

KW - Slow freezing

KW - Vitrification

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