Dadashpour Davachi Navid, Zare Shahneh Ahmad, Kohram Hamid, Zhandi Mahdi, Dashti Saeed, Shamsi Helia, Moghadam Razieh
Department of Animal Science, Faculty College of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran.
Department of Animal Science, Faculty College of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran ; Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, IR Iran.
Iran Red Crescent Med J. 2014 Sep 5;16(9):e20749. doi: 10.5812/ircmj.20749. eCollection 2014 Sep.
To current knowledge, different oocyte's recovery method and various seasons have profound impact on in vitro embryo production (IVEP).
The aim of this study was to define an efficient recovery method for oocytes harvesting from slaughterhouse material in different seasons, and their effects on IVEP yield.
Ovaries from slaughtered ewes in breeding season (BS) and non-breeding season (NBS) were collected from a local abattoir. The oocytes were recovered through aspiration, centrifugation (ORC), puncture and slicing, and categorized into three classes (I, oocytes with more than three layers of cumulus cells; II, less than three layers with damaged cumulus cells; III, denuded oocytes). After cultivation in TCM 199 for 24 hours, matured oocytes were subjected to in vitro fertilization (IVF) and in vitro culture (IVC). The oocyte recovery using ORC in BS and NBS was significantly higher (P < 0.05) compared with other recovery methods.
No significant dissimilarities in the proportion of oocytes reaching M-II stage were recorded when using different oocyte recovery methods in different seasons. Aspiration resulted in lower (P < 0.05) proportion of class I (BS, 60.0 ± 2.1; NBS, 51.1 ± 2.1) compared to ORC (BS, 82.0 ± 1.2; NBS, 70.0 ± 1.2), slicing (BS, 80.0 ± 2.1; NBS, 71.0 ± 1.4) and puncture (BS, 80.0 ± 1.5; NBS, 72.0 ± 2.0). Monospermy and blastocyst development rates were significantly higher using ORC than other recovery techniques in both BS and NBS. More oocytes with high quality, greater blastocyst development and oocyte recovery rates were achieved in BS.
The results revealed that oocytes harvesting technique and season are effective in the rate of cleavage and blastocysts' development, and suggest that despite same meiotic resumption rate in all treatments, it would be better to use ORC.
就目前所知，不同的卵母细胞回收方法和不同季节对体外胚胎生产（IVEP）有深远影响。
本研究的目的是确定一种在不同季节从屠宰场材料中采集卵母细胞的有效回收方法，以及它们对IVEP产量的影响。
从当地屠宰场收集繁殖季节（BS）和非繁殖季节（NBS）屠宰母羊的卵巢。通过抽吸、离心（ORC）、穿刺和切片回收卵母细胞，并分为三类（I，有三层以上卵丘细胞的卵母细胞；II，少于三层且卵丘细胞受损的卵母细胞；III，裸卵）。在TCM 199中培养24小时后，将成熟卵母细胞进行体外受精（IVF）和体外培养（IVC）。与其他回收方法相比，在BS和NBS中使用ORC回收的卵母细胞显著更高（P < 0.05）。
在不同季节使用不同的卵母细胞回收方法时，达到M-II期的卵母细胞比例没有显著差异。与ORC（BS，82.0 ± 1.2；NBS，70.0 ± 1.2）、切片（BS，80.0 ± 2.1；NBS，71.0 ± 1.4）和穿刺（BS，80.0 ± 1.5；NBS，72.0 ± 2.0）相比，抽吸导致I类卵母细胞比例较低（P < 0.05）（BS，60.0 ± 2.1；NBS，51.1 ± 2.1）。在BS和NBS中，使用ORC的单精子受精和囊胚发育率均显著高于其他回收技术。在BS中获得了更多高质量的卵母细胞、更高的囊胚发育率和卵母细胞回收率。
结果表明，卵母细胞采集技术和季节对卵裂率和囊胚发育率有影响，并且表明尽管所有处理中的减数分裂恢复率相同，但使用ORC会更好。
