In vitro maturation and transmission electron microscopic observation of horse oocytes after vitrification. 1996

S Hochi, and M Kozawa, and T Fujimoto, and E Hondo, and J Yamada, and N Oguri
Laboratory of Horse Production, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.

The study was designed to examine the suitability of immature horse oocytes for vitrification. Immature oocytes derived from slaughtered horse ovaries were transferred to a vitrification solution (EFS; 40% ethylene glycol, 18% Ficoll, and 0.3 M sucrose in modified phosphate-buffered saline) directly (Groups 1 and 4) or were first exposed to 20% ethylene glycol solution for 10 min (Groups 2 and 5) or 20 min (Groups 3 and 6). Oocytes were handled at 20 degrees C (Groups 1, 2, and 3) or 30 degrees C (Groups 4, 5, and 6). After vitrification and warming, their viability was assessed by maturation culture for 32 h. The percentages of oocytes reaching the metaphase II stage after the in vitro maturation in Groups 2, 3, 5, and 6 (16.0, 16.7, 10.0. and 8.2%, respectively) were higher than those in Groups 1 and 4 (2.2 and 1.9%, respectively). In untreated control oocytes, 55.6% completed meiosis in vitro. Transmission electron microscopy was used to compare the fine structure of vitrified oocytes (treated as Group 2) with those of untreated control oocytes and EFS-exposed, nonvitrified oocytes (n = 10 each). The viability of EFS-exposed oocytes, assessed by in vitro maturation, was 27.7%. Vitrification induced some ultrastructural changes, such as the swelling of mitochondria together with reduced matrix density, the destruction of communication between oocytes and their surrounding cumulus cells, and the presence of vacuoles located in the periphery of the ooplasm. However, these changes were not always observed. Exposure of the oocytes to EFS solution induced similar ultrastructural changes in mitochondria and cell-cell communication but to a lesser extent. However, the exposure to EFS induced vacuoles in the periphery of the ooplasm to the same extent as did the vitrification. Thus, immature horse oocytes can be cryopreserved by vitrification with EFS solution. Reduced viability of EFS-exposed and/or vitrified horse oocytes may relate to morphological changes such as destruction of the intercellular communications between cumulus cells and oocytes.

UI MeSH Term Description Entries
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D009865 Oocytes Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM). Ovocytes,Oocyte,Ovocyte
D002470 Cell Survival The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability. Cell Viability,Cell Viabilities,Survival, Cell,Viabilities, Cell,Viability, Cell
D003451 Cryoprotective Agents Substances that provide protection against the harmful effects of freezing temperatures. Cryoprotective Agent,Cryoprotective Effect,Cryoprotective Effects,Agent, Cryoprotective,Agents, Cryoprotective,Effect, Cryoprotective,Effects, Cryoprotective
D005026 Ethylene Glycols An ethylene compound with two hydroxy groups (-OH) located on adjacent carbons. They are viscous and colorless liquids. Some are used as anesthetics or hypnotics. However, the class is best known for their use as a coolant or antifreeze. Dihydroxyethanes,Ethanediols,Glycols, Ethylene
D005069 Evaluation Studies as Topic Works about studies that determine the effectiveness or value of processes, personnel, and equipment, or the material on conducting such studies. Critique,Evaluation Indexes,Evaluation Methodology,Evaluation Report,Evaluation Research,Methodology, Evaluation,Pre-Post Tests,Qualitative Evaluation,Quantitative Evaluation,Theoretical Effectiveness,Use-Effectiveness,Critiques,Effectiveness, Theoretical,Evaluation Methodologies,Evaluation Reports,Evaluation, Qualitative,Evaluation, Quantitative,Evaluations, Qualitative,Evaluations, Quantitative,Indexes, Evaluation,Methodologies, Evaluation,Pre Post Tests,Pre-Post Test,Qualitative Evaluations,Quantitative Evaluations,Report, Evaluation,Reports, Evaluation,Research, Evaluation,Test, Pre-Post,Tests, Pre-Post,Use Effectiveness
D005260 Female Females
D005362 Ficoll A sucrose polymer of high molecular weight.
D006736 Horses Large, hoofed mammals of the family EQUIDAE. Horses are active day and night with most of the day spent seeking and consuming food. Feeding peaks occur in the early morning and late afternoon, and there are several daily periods of rest. Equus caballus,Equus przewalskii,Horse, Domestic,Domestic Horse,Domestic Horses,Horse,Horses, Domestic
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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