In vitro maturation of equine oocytes collected by follicle aspiration and by the slicing of ovaries.
Abstract: The aim of this study was to examine 2 techniques for oocyte recovery from equine ovaries at slaughter: by aspiration of follicles and by additional slicing of ovaries. The morphology and nuclear configuration of oocytes recovered with either technique, and the time course of nuclear maturation during in vitro maturation were evaluated. Recovery rates were 1.75 and 4.14 oocytes per ovary for aspiration and slicing (total 145 and 344 oocytes from 83 ovaries), respectively. The oocytes were classified according to their cumulus/ooplasm morphology into 4 groups: compact/circular(A), compact/semicircular(B), expanded(C) and others(D). The percentages of oocytes in Groups A, B, C and D were 34, 38, 25 and 3% (aspiration) and 55, 26, 17 and 3% (slicing), respectively. The proportions of oocytes with a germinal vesicle in Groups A, B, C and D were 28 29 (97%), 23 35 (66%), 11 23 (48%) and 2 4 (50%) in oocytes from aspiration and 91 100 (91%), 52 65 (80%), 15 29 (52%) and 1 2 (50%) in oocytes from slicing, respectively. Group A and B oocytes recovered by aspiration (n=212) and slicing (n=312) were cultured in TCM199 supplemented with 10% fetal bovine serum, 1 mug/ml estradiol-17beta, and 0.02 AU/ml FSH at 38.5 degrees C in 5% CO(2) in air (5 to 10 oocytes per 50- mu l microdrop). At 8, 16, 24, 32 and 40 h of culture, the oocytes were fixed and stained. There were no significant differences in the percentages of Metaphase II stage (MII)-oocytes between recovery techniques at any time points examined. The proportions of MII-oocytes were 1 42 (2%), 4 43 (9%), 21 42 (50%), 28 45 (62%), and 28 40 (70%) at the respective time point in oocytes from aspiration and 0 51 (0%), 3 54 (6%), 22 59 (37%), 43 72 (60%), and 51 76 (67%) in oocytes from slicing, respectively. In most of the oocytes, resumption of meiosis occurred between 8 and 16 h of culture. The proportions of MII-oocytes increased significantly between 16 and 24 h and between 24 and 32 h of culture.
Publication Date: 1993-11-01 PubMed ID: 16727378DOI: 10.1016/0093-691x(93)90364-bGoogle Scholar: Lookup
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Summary
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This research focused on comparing two methods of oocyte recovery in horse ovaries at slaughter, namely aspiration and slicing. The researchers were interested in the morphology, nuclear configuration, and timeline of the maturation process of the oocytes collected. No significant differences were found in the maturation status of oocytes between the two recovery techniques.
Oocyte Recovery Methods: Aspiration and Slicing
- The researchers examined the efficacy of two different techniques for recovering oocytes: one involving aspiration of follicles, and another involving the slicing of the ovaries.
- The recovery rates were 1.75 and 4.14 oocytes per ovary for aspiration and slicing respectively, showing a more substantial recovery with the slicing technique. A total of 145 oocytes were recovered via aspiration, and 344 oocytes by slicing from 83 ovaries.
Oocyte Classification by Morphology
- The oocytes were further classified into four groups based on their cumulus/ooplasm morphology: compact/circular (Group A), compact/semicircular (Group B), expanded (Group C), and others (Group D).
- Group A had the highest percentage of oocytes in slicing (55%) followed by aspiration method (34%). For Group B, the percentages were about balanced between slicing and aspiration. For Group C and D, the percentages were relatively low, under 25% regardless of the method of recovery.
Nuclear Configuration of Oocytes and Maturation Process
- The proportion of oocytes containing a germinal vesicle, an indicator of immaturity, was determined in each group. A significantly higher percentage of these immature oocytes were found in Groups A and B, regardless of the method of recovery.
- Over time, the number of oocytes reaching the Metaphase II stage (MII) – the final stage of maturation – was tracked. Oocytes recovered both by aspiration and by slicing started meiosis between 8 and 16 hours of culture, and the proportion of MII-stage oocytes increased notably between 16 and 24 hours and between 24 and 32 hours of culture.
- Overall, there were no significant differences in the maturation percentages of MII-oocytes between the aspiration and slicing techniques at any time points examined. Therefore, both techniques proved to be equally suitable for oocyte recovery in the studied context.
Cite This Article
APA
Choi YH, Hochi S, Braun J, Sato K, Oguri N.
(1993).
In vitro maturation of equine oocytes collected by follicle aspiration and by the slicing of ovaries.
Theriogenology, 40(5), 959-966.
https://doi.org/10.1016/0093-691x(93)90364-b Publication
Researcher Affiliations
- Laboratory of Horse Production Obihiro, Hokkaido 080, Japan.
Citations
This article has been cited 3 times.- Bahrami M, Morris MB, Day ML. Amino acid supplementation of a simple inorganic salt solution supports efficient in vitro maturation (IVM) of bovine oocytes. Sci Rep 2019 Aug 13;9(1):11739.
- Franciosi F, Tessaro I, Dalbies-Tran R, Douet C, Reigner F, Deleuze S, Papillier P, Miclea I, Lodde V, Luciano AM, Goudet G. Analysis of Chromosome Segregation, Histone Acetylation, and Spindle Morphology in Horse Oocytes. J Vis Exp 2017 May 11;(123).
- Abdoon AS, Abdel-Rahman HA, Shawki SM, Kandil OM, Fathalla SI. Influence of follicle size, methods of retrieval on oocytes yield and morphology in Egyptian Jennies ovaries with special reference to maturation rate in vitro. Vet Res Commun 2014 Dec;38(4):287-95.
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