Alvarez R H, Meneghel M, Martinez A C, Pires Rml, Schammass E A
Genetic and Animal Reproduction Center, Instituto de Zootecnia - APTA, Nova Odessa-SP, Brazil.
Reprod Domest Anim. 2008 Jun;43(3):257-260. doi: 10.1111/j.1439-0531.2007.00884.x. Epub 2007 Dec 30.
The aim of this study was to evaluate if blastocysts arising from in vitro culture of Grade 3 bovine morulae produced in vivo can promote acceptable pregnancy rates when transferred into recipients. Embryos of different stages and qualities were recovered from superovulated Bos taurus and B. indicus donors. Grade 3 morulae were cultured in either Holding Plus or TCM-199 (supplemented with 10% bovine fetal serum) media for 24 h at 38.5 degrees C. After this culture period, the resulting blastocysts were morphologically classified (Grades 1, 2 and 3) and transferred into recipients previously synchronized with the donors. Non-cultured Grades 1 and 3 morulae were used as control. Pregnancy diagnosis was carried out 60 days after embryo transfer and the data were analysed by logistic regression, considering variables, such as embryo quality (Grade), donor breed, culture medium, donor-recipient synchrony and seasonality. Embryo quality was the only variable, showing significant effect on the pregnancy rate. Pregnancy rates for non-cultured Grade 1 and 3 morulae, and blastocysts arising from cultured Grade 3 morulae were 58.1% (n = 31), 17.1% (n = 35) and 51.1% (n = 47), respectively (p < 0.05). There were no statistical differences between non-cultured Grade 1 morulae and cultured blastocysts. Pregnancy rates for Grades 1 (65.0%) and 2 (60.0%) were higher than Grade 3 (29.4%) cultured blastocysts (p < 0.05). It was concluded that short-term in vitro culture is a very convenient method of identifying morphologically low quality morulae with higher chances of continuing development after the transfer into recipients.
本研究的目的是评估体内产生的3级牛桑葚胚经体外培养形成的囊胚移植到受体后能否获得可接受的妊娠率。从超排的黄牛和印度瘤牛供体中回收不同阶段和质量的胚胎。将3级桑葚胚在 Holding Plus 或 TCM - 199(添加10%牛胎牛血清)培养基中于38.5℃培养24小时。培养期结束后,对所得囊胚进行形态学分级(1级、2级和3级),并移植到先前与供体同步的受体中。未培养的1级和3级桑葚胚用作对照。胚胎移植60天后进行妊娠诊断,并通过逻辑回归分析数据,考虑胚胎质量(等级)、供体品种、培养基、供体 - 受体同步性和季节性等变量。胚胎质量是唯一对妊娠率有显著影响的变量。未培养的1级和3级桑葚胚以及由培养的3级桑葚胚形成的囊胚的妊娠率分别为58.1%(n = 31)、17.1%(n = 35)和51.1%(n = 47)(p < 0.05)。未培养的1级桑葚胚与培养的囊胚之间无统计学差异。1级(65.0%)和2级(60.0%)培养囊胚的妊娠率高于3级(29.4%)培养囊胚(p < 0.05)。得出的结论是,短期体外培养是一种非常便捷的方法,可用于识别形态学上质量较低但移植到受体后继续发育机会较高的桑葚胚。