A Comparative Study on Meiotic Competence of Pig Oocyte by Two Methods for Collection: Aspiration and Dissection
The aim of this study is to examine the effect of different collection technique to the developmental meiotic competent after in vitro maturation (IVM). Ovaries were collected from local adult and non-pregnant slaughtered pig. Porcine oocytes were collect
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Abstract
The aim of this study is to examine the effect of different collection technique to the developmental meiotic competent after in vitro maturation (IVM). Ovaries were collected from local adult and non-pregnant slaughtered pig. Porcine oocytes were collected by both aspiration method and dissection technique. Follicle isolated from dissection are classified into 3 groups: Group 1: 3–4 mm, Group 2: 4–5 mm, Group 3: 5–6 mm. Oocytes from aspiration technique take place as another group. These groups were cultured in maturation medium for 40–42 h, at 38.5 °C, 5% CO2. After maturation process, the meiotic competence of oocytes was examined by orcein staining. The results show that the percentages of oocyte reach to metaphase II from Group 3 (68.19%), Group 2 (52.12%) and Aspiration group (39.2%) are significantly higher than Group 1 (7.15%), especially, Group 3 is significantly higher than the Aspiration group. Therefore, it is the most effective way to use oocytes from follicles of 4–6 mm in diameter, especially 5–6 mm, for IVM process in experimental research and subsequent technology. We suggest that the oocytes from Group 1 (follicle size 3–4 mm) are needed to pass a pre-IVM or incubate in IVG for a short time before introducing to IVM process. Keywords
Porcine oocyte
1
Antral follicle
Introduction
In nature, an ovary has a variety of follicles containing oocytes. Before the ovulation, oocyte maturation is generally illustrated as the process from when signals initiate germinal vesicle breakdown to a completion of the nuclear changes resulting in the expulsion of the first polar body [1]. As the follicle increases in size, the oocyte from within it is more likely to be fertilized, cleave, form a blastocyst and implant [2]. Thus, a majority of people often use follicles that have the sizes of 3–6 mm in diameter to isolate oocytes for culture H.K. Dang P.T. Duy L.T.H. Linh N. Van Thuan H-T. Bui (&) Cellular Reprogramming Laboratory, School of Biotechnology, International University, Vietnam National University, 6 Linh Trung, Thu Duc District, Ho Chi Minh City, 70000, Vietnam e-mail: [email protected]
Aspiration
Dissection
In vitro maturation
in vitro maturation (IVM) by aspiration or dissection method [3–5]. However, only pig oocytes in antral follicles of 4– 6 mm with 122.1 ± 2.2 lm diameter, acquire the meiotic competence because they are approximately approached at the fully grown oocytes stages [6]. Due to the oocytes are very sensitive and easily affected by outer force, it is important to decide the way of isolating them from the follicles. In this case, the dissection method is preferred rather than the aspiration to collect the oocytes. Using dissection offer some advantages that aspiration could not do. At first, dissection technique required us to use medical blades for cutting the ovarian cortical and tissue as well as breaking the follicle membrane. This work does not impact the oocyte because it is still kept in the follicle, without applying any force to them. With aspi
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