Cocero M J, Alabart J L, Hammami S, Martí J I, Lahoz B, Sánchez P, Echegoyen E, Beckers J F, Folch J
Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.
Reprod Domest Anim. 2011 Jun;46(3):463-70. doi: 10.1111/j.1439-0531.2010.01690.x. Epub 2010 Oct 1.
In vitro oocyte maturation can be influenced by oocyte source and maturation media composition. The aim of the present study was to compare the efficiency of a defined in vitro maturation medium (TCM199 supplemented with cysteamine and epidermal growth factor; Cys + EGF) with an undefined medium (TCM199 supplemented with follicle-stimulating hormone and follicular fluid; FSH + FF) for in vitro production (IVP) of ovine embryos, using oocytes obtained by laparoscopic ovum pick-up from FSH-stimulated [n=11; 158 cumulus-oocyte complexes (COCs)] and non-stimulated (n=16; 120 COCs) live ewes, as well as abattoir-derived oocytes (170 COCs). The produced blastocysts were vitrified and some of them were transferred to synchronized recipients. The best and the worst final yields of embryo IVP observed in this study were obtained using oocytes from FSH-stimulated ewes matured in FSH + FF (41.3%; 33/80) and in Cys + EGF (19.2%; 15/78) medium, respectively (p<0.01). No significant differences between both media were attained in the blastocyst development rate or in the final yield of embryo IVP using oocytes from non-stimulated ewes or abattoir-derived oocytes. The overall in vivo survival rate of the transferred vitrified blastocysts was 13.1% (8/61), without significant differences between oocyte sources or maturation media. In conclusion, under the experimental conditions of the present study, TCM199 supplemented with cysteamine and EGF is a convenient defined maturation medium for IVP of embryos from oocytes of live non-stimulated ewes or from oocytes of abattoir-derived ovaries. However, the best final yield of embryo IVP observed in this study was attained when oocytes came from FSH-stimulated donors and TCM199 was supplemented with FSH and follicular fluid.
体外卵母细胞成熟可受卵母细胞来源和成熟培养基成分的影响。本研究的目的是比较一种成分明确的体外成熟培养基(添加半胱胺和表皮生长因子的TCM199;Cys + EGF)与一种成分不明确的培养基(添加促卵泡激素和卵泡液的TCM199;FSH + FF)用于绵羊胚胎体外生产(IVP)的效率,所使用的卵母细胞通过腹腔镜采卵从经促卵泡激素刺激的母羊(n = 11;158个卵丘 - 卵母细胞复合体[COCs])和未经刺激的母羊(n = 16;120个COCs)以及屠宰场来源的卵母细胞(170个COCs)中获取。所产生的囊胚经玻璃化处理,其中一些被移植到同期受体中。本研究中观察到的胚胎IVP最佳和最差最终产率分别是使用在FSH + FF培养基(41.3%;33/80)和Cys + EGF培养基(19.2%;15/78)中成熟的经促卵泡激素刺激的母羊的卵母细胞获得的(p<0.01)。使用未经刺激的母羊或屠宰场来源的卵母细胞时,两种培养基在囊胚发育率或胚胎IVP最终产率方面均未达到显著差异。移植的玻璃化囊胚的总体体内存活率为13.1%(8/61),卵母细胞来源或成熟培养基之间无显著差异。总之,在本研究的实验条件下,添加半胱胺和表皮生长因子的TCM199是用于从未经刺激的活体母羊的卵母细胞或屠宰场来源的卵巢卵母细胞进行胚胎IVP的一种方便的成分明确的成熟培养基。然而,本研究中观察到的胚胎IVP最佳最终产率是当卵母细胞来自经促卵泡激素刺激的供体且TCM199添加促卵泡激素和卵泡液时获得的。