Reproductive Phases Influence on In Vitro Maturation of Abattoir-Derived Bovine Oocytes

Z. C. Das, M. Rashid, M. Hoque, M. Haider, A. Talukder, A. Rahman
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Abstract

This study investigated the influence of the reproductive phase of oocyte donors originating from abattoir-derived ovaries on in vitro maturation (IVM) of bovine oocytes. The IVM of the oocytes was evaluated by observing the degree of cumulus cell (CC) expansion and extrusion of the first polar body (PB-1) as well as both nuclear progression (NP) and cytoplasmic maturation (CM).  For NP, after 24 h of IVM, oocytes were denuded and stained with DNA fluorochrome and observed under an epifluorescence microscope. The male or female pronucleus (PN) forming ability of an oocyte following in vitro fertilization (IVF) was considered as CM. For CM, the fertilized zygotes after 24 h post-insemination were stained with DNA fluorochrome and observed under an epifluorescence microscope. Results revealed that the maturation rate was significantly influenced by the reproductive phase of the oocytes donor, luteal phase ovaries (LPO) versus follicular phase ovaries (FPO) based on the presence or absence of corpus luteum (CL), respectively. The significantly higher CC expansion (2°; 44.9±4.9% versus 35.9±2.7%) and PB-1 extrusion (58.3±1.7% versus 46.7±2.4%) were observed with LPO than FPO oocytes, respectively. A significant (p<0.05) effect of the reproductive phase was also noted with LPO oocytes progressing higher rate of mature metaphase II stage (MII; 58.3±1.7% versus 46.7±2.4%) and PN (54.9±2.3% versus 44.4±3.0%) formation following IVF than FPO oocytes that imply better NP and CM of oocytes, respectively. Therefore, the present study indicates that the reproductive phase of oocyte donors notably from LPO positively influences the IVM of bovine oocytes to produce a higher number of viable embryos in vitro. Ann. Bangladesh Agric. (2022) 26 (1) : 1-19
生殖阶段对牛源性卵母细胞体外成熟的影响
本研究研究了来自屠宰场的卵巢的卵母细胞供体的生殖阶段对牛卵母细胞体外成熟(IVM)的影响。通过观察卵母细胞第一极体(PB-1)的积云细胞(cumulus cell, CC)扩张和挤压程度以及核进展(NP)和细胞质成熟(cytoplasmic maturity, CM)来评价卵母细胞的IVM。对于NP,在IVM 24 h后,剥去卵母细胞,用DNA荧光染色,在荧光显微镜下观察。体外受精(IVF)后卵母细胞形成雄性或雌性原核(PN)的能力被认为是CM。对CM,在授精24 h后的受精卵进行DNA荧光染色,在荧光显微镜下观察。结果表明,卵母细胞供体的生殖期、黄体期卵巢(LPO)和卵泡期卵巢(FPO)在黄体存在或不存在的基础上对成熟率有显著影响。CC膨胀率显著提高(2°;LPO比FPO分别为44.9±4.9%(35.9±2.7%)和58.3±1.7%(46.7±2.4%)。生殖期也有显著影响(p<0.05), LPO卵母细胞在成熟中期(MII;(58.3±1.7%对46.7±2.4%)和PN(54.9±2.3%对44.4±3.0%)的形成均优于FPO卵母细胞,表明卵母细胞的NP和CM分别更好。因此,本研究表明,特别是来自LPO的卵母细胞供体的生殖期对牛卵母细胞的IVM产生积极的影响,从而在体外产生更多的活胚。孟加拉国阿格利司。(2022) 26 (1): 1-19
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