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Addition of ascorbate during cryopreservation stimulates subsequent embryo development.
Michelle Lane1, Jeffery M Maybach, David K Gardner
1Research Department, Colorado Center for Reproductive Medicine, 799 East Hampden Ave, Suite 520, Englewood 80110, USA. mlane@colocrm.com
Human Reproduction (Oxford, England)
|September 28, 2002
Summary
Adding the antioxidant ascorbate to cryopreservation solutions significantly improves mammalian embryo development by reducing oxidative stress. Ascorbate enhances inner cell mass development and metabolism, especially in slow-frozen embryos.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cryobiology
Background:
- Cryopreservation of mammalian embryos reduces subsequent development compared to fresh embryos.
- Cryopreservation induces oxidative stress in embryos, evidenced by increased hydrogen peroxide levels.
- Antioxidants may mitigate cryopreservation-induced damage to embryos.
Purpose of the Study:
- To investigate the impact of adding the antioxidant ascorbate to cryopreservation solutions on embryo development.
- To assess the effect of ascorbate on oxidative stress markers and embryo physiology post-cryopreservation.
Main Methods:
- Mouse embryos (2-cell and blastocyst stages) were cryopreserved using slow-freezing or vitrification.
- Cryopreservation solutions contained either no ascorbate or 0.1 or 0.5 mmol/l ascorbate.
- Assessed hydrogen peroxide levels, inner cell mass development, lactate dehydrogenase leakage, and metabolic rates.
Main Results:
- Ascorbate addition reduced hydrogen peroxide levels in cryopreserved embryos.
- 0.1 mmol/l ascorbate significantly enhanced inner cell mass development in blastocysts.
- Ascorbate-treated embryos exhibited reduced lactate dehydrogenase leakage and increased metabolism, with greater benefits observed in slow-frozen embryos.
Conclusions:
- Ascorbate supplementation in cryopreservation solutions is valuable for improving mammalian embryo development.
- 0.1 mmol/l ascorbate is a beneficial concentration for cryopreservation protocols.
- This strategy effectively combats oxidative stress during cryopreservation.