Hammami S, Morató R, Romaguera R, Roura M, Catalá M G, Paramio M T, Mogas T, Izquierdo D
Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Reprod Domest Anim. 2013 Apr;48(2):339-44. doi: 10.1111/j.1439-0531.2012.02160.x. Epub 2012 Aug 22.
The aim of this study was to test the effect of insulin-transferrin-selenium (ITS) and L-ascorbic acid (AA) supplementation and the hormonal level during in vitro maturation (IVM) of small oocytes from pre-pubertal goat on the blastocyst yield and quality. Concretely, we used four maturation media: conventional IVM medium (CM), growth medium (GM: CM+ITS+AA and low level of hormones), modified CM (mCM: CM with low level of hormones) and modified GM (mGM: CM+ITS+AA and normal level of hormones). Cumulus-oocyte complexes (COCs) were classified into two categories according to oocyte diameter: <125 μm and ≥ 125 μm. Large oocytes were matured 24 h in CM (Treatment A). Small oocytes were matured randomly in six experimental groups: Treatment B: 24 h in CM; Treatment C: 12 h in GM and 12 h in CM; Treatment D: 24 h in mGM; Treatment E: 12 h in mGM and 12 h in CM; Treatment F: 12 h in mCM and 12 h in CM; and Treatment G: 12 h in GM and 12 h in mGM. After IVM, oocytes were fertilized and cultured for 8 days. The blastocyst quality was assessed by the survival following vitrification/warming and the mean cell number. When different maturation media were combined, the blastocyst rate did not improve. The large oocytes produced the highest blastocysts yield. However, the culture of small oocytes in GM (53.3%) enhanced the post-warming survival of blastocysts compared to large oocytes matured in CM (35.7%). In conclusion, IVM of pre-pubertal goat small oocytes in GM would be useful to improve the quality of in vitro-produced blastocysts.
本研究旨在测试补充胰岛素-转铁蛋白-硒(ITS)和L-抗坏血酸(AA)以及青春期前山羊小卵母细胞体外成熟(IVM)期间的激素水平对囊胚产量和质量的影响。具体而言,我们使用了四种成熟培养基:常规IVM培养基(CM)、生长培养基(GM:CM + ITS + AA和低水平激素)、改良CM(mCM:含低水平激素的CM)和改良GM(mGM:CM + ITS + AA和正常水平激素)。卵丘-卵母细胞复合体(COCs)根据卵母细胞直径分为两类:<125μm和≥125μm。大卵母细胞在CM中成熟24小时(处理A)。小卵母细胞随机分为六个实验组:处理B:在CM中成熟24小时;处理C:在GM中成熟12小时,然后在CM中成熟12小时;处理D:在mGM中成熟24小时;处理E:在mGM中成熟12小时,然后在CM中成熟12小时;处理F:在mCM中成熟12小时,然后在CM中成熟12小时;处理G:在GM中成熟12小时,然后在mGM中成熟12小时。IVM后,卵母细胞受精并培养8天。通过玻璃化/复温后的存活率和平均细胞数评估囊胚质量。当不同的成熟培养基组合使用时,囊胚率并未提高。大卵母细胞产生的囊胚产量最高。然而,与在CM中成熟的大卵母细胞(35.7%)相比,在GM中培养的小卵母细胞(53.3%)提高了复温后囊胚的存活率。总之,在GM中对青春期前山羊小卵母细胞进行IVM有助于提高体外生产囊胚的质量。