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. 2020 Apr 10;10(4):658.
doi: 10.3390/ani10040658.

L-OPU in Goat and Sheep-Different Variants of the Oocyte Recovery Method

Jarosław Wieczorek  1 , Jurij Koseniuk  2 , Maria Skrzyszowska  3 , Mirosław Cegła  3
Affiliations

L-OPU in Goat and Sheep-Different Variants of the Oocyte Recovery Method

Jarosław Wieczorek et al. Animals (Basel). .

Abstract

The laparoscopic method of recovering oocytes in goats and sheep is one of the minimally invasive methods used in the biotechnology of animal reproduction. It allows for good quality oocytes that are suitable for in vitro maturation and fertilization to be recovered. The limitation of using the laparoscopic ovum pick-up (L-OPU) method in goat and sheep is its changing effectiveness and the lack of repeatability of results, as well as the varying effectiveness of different variants of the method. Therefore, it is necessary to develop effective non-invasive techniques allowing for multiple good quality oocyte recovery that would be suitable for in vitro maturation and fertilization. In this study, four different L-OPU variants were described in goats and sheep. Various techniques of recovering oocytes were discussed, including the techniques of conducting the operation, various tools for recovering oocytes, and different plans of hormonal stimulation. Recovery rates were 35% (Variant I), 57% (Variant II), 72% (Variant III), and 67% (Variant IV). After evaluation, 94% (both Variant I and II), 93% (Variant III), and 84% (Variant IV) of the oocytes were qualified for in vitro maturation. The results of the study show that the proposed technique of laparoscopic recovery of oocytes allows a sufficient number of ovarian cells suitable for in vitro culture to be obtained and as a consequence it makes them useful in in vitro maturation/in vitro fertilization (IVM/IVF) programs or cloning. The method allows for a fast and effective conduct of the operation in a living donor with minimal invasiveness while preserving the excellent condition of animals.

Keywords: goat; laparoscopy; oocyte; ovum pick up; sheep.

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Conflict of interest statement

The authors declare that they have no conflicts of interest to disclose.

Figures

Figure 1
Figure 1
Schedule of estrus synchronization with Cronolone and superovulation with pregnant mare serum gonadotropin (PMSG) or follicle-stimulating hormone (FSH) treatment.
Figure 2
Figure 2
Oocyte recovery—aspiration of fluid from ovarian follicles with oocytes. (A)—follicle before aspiration, (B)—follicle after aspiration.
Figure 3
Figure 3
Tools for oocyte aspiration. A—originally designed 1 mL syringe for oocyte aspiration with a needle (needle: 1 cm long and 0.6mm diameter, 23 G; Polish Patent Application P-361068, 2003) (Variants I and II), B—catheter for oocyte aspiration (Variants III and IV).
Figure 4
Figure 4
Immature oocytes collected using the ovum pick-up technique. (A) Class I—excellent quality of cumulus–oocyte complexes (COCs); morphologically normal oocytes with evenly granulated (homogenous) cytoplasm, surrounded by thick compact layers of cumulus cells; (B) class II—good quality COCs, morphologically normal oocytes with evenly granulated (homogenous) cytoplasm, surrounded by at least three compact layers of cumulus cells; (C) class III—cumulus cell-deprived oocytes (naked oocytes); (D) class IV—poor quality oocytes with clearly visible degenerative (necrotic or apoptotic) changes in cytoplasm.
Figure 5
Figure 5
Comparing various L-OPU sessions with respect to percentages of recovered oocytes and oocytes selected for in vitro maturation (IVM) in sheep. (Variant I—sessions 1–3, Variant II—sessions 4–6, Variant III—sessions 7–13, Variant IV—sessions 14–17).
Figure 6
Figure 6
Timing of the L-OPU procedure in goats.
Figure 7
Figure 7
Comparing various L-OPU sessions with respect to percentages of recovered oocytes and oocytes selected for IVM in sheep.
Figure 8
Figure 8
Timing of the L-OPU procedure in sheep in consecutive sessions.
See this image and copyright information in PMC

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