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Porcine oocyte vitrification in optimized low toxicity solution with open pulled straws.
F Marco-Jiménez1, L Casares-Crespo2, J S Vicente2
1Laboratory of Biotechnology of Reproduction, Institute of Science and Animal Technology (ICTA) at the Polytechnic University of Valencia, C/Camino de Vera s/n, 46022 Valencia, Spain.
Optimizing the VM3 cryoprotectant solution to 5.6 M enabled efficient vitrification of porcine oocytes using the open pulled straw device. While survival rates were maintained, chromosomal organization and cortical granule distribution were negatively impacted by the vitrification process.
Area of Science:
- Reproductive biology
- Cryobiology
- Oocyte cryopreservation
Background:
- Developing efficient oocyte cryopreservation methods is crucial for both human and domestic animal reproduction.
- The VM3 solution is a low-toxicity cryoprotectant, but its optimal concentration for vitrification needs refinement.
Purpose of the Study:
- To optimize the VM3 solution for vitrifying porcine oocytes using the open pulled straw (OPS) device.
- To assess the effects of optimized VM3 solution on oocyte viability, chromosomal organization, and cortical granule distribution.
Main Methods:
- Determined the minimum effective cryoprotectant concentration of VM3 for vitrification using OPS.
- Examined oocyte ultrastructure via cryo-scanning electron microscopy.
- Assessed oocyte viability, chromosomal organization, and cortical granule distribution using fluorescent staining after warming.
Main Results:
- The minimum VM3 concentration for successful vitrification with OPS was 5.6 M.
- Oocyte survival rates post-warming were comparable to fresh oocytes.
- Vitrification significantly impaired chromosomal organization and cortical granule distribution, irrespective of the VM3 solution used.
Conclusions:
- VM3 solution can be optimized to 5.6 M for porcine oocyte vitrification with OPS.
- The optimized VM3 solution did not improve chromosomal organization or cortical granule distribution compared to the original VM3.
- The vitrification process itself, rather than the VM3 concentration, significantly affected oocyte quality post-warming.
