van Wagtendonk-de Leeuw A M, Mullaart E, de Roos A P, Merton J S, den Daas J H, Kemp B, de Ruigh L
Holland Genetics, R&D, Arnhem, The Netherlands.
Theriogenology. 2000 Jan 15;53(2):575-97. doi: 10.1016/s0093-691x(99)00259-9.
Since the introduction of in vitro production (IVP) of bovine and sheep pre-implantation embryos, increased birth weights and other deviations of IVP calves and lambs compared with AI or MOET offspring have been reported. Study 1 of the present paper, a comparison between AI, MOET and IVP (co-culture/serum) calves with respect to calf and calving characteristics in large-scale field conditions, confirms these reports. In addition, it is shown that MOET calves tend towards higher birth weights and have significantly longer gestations and more difficult calvings than AI calves. It is presently unknown if the effect of IVP is also observed later in life. In this paper, data on reproduction characteristics of bovine IVP co-culture/serum offspring are presented. Semen production--and non return data of one year old IVP bulls and superovulation-, AI- and OPU/IVP results of one year old IVP heifers are compared with those of one year old AI and MOET animals producing semen or embryos in the same time period. So far, there are no indications that the use of IVP is reflected in deviate reproduction characteristics of bovine IVP offspring. It has been suggested that use of co-culture cells and serum during in vitro culture of bovine (and sheep) embryos may partially explain the increased birth weights and other deviations of bovine and sheep IVP offspring. Deletion of these factors in semi-defined culture media, e.g. Synthetic Oviductal Fluid (SOF), could result in more normal offspring. Study 2 investigates this hypothesis in both field conditions (Study 2a, comparing AI, IVP co-culture/serum and IVP SOF calves) and in semi-standardized conditions (Study 2b, comparing MOET, IVP co-culture/serum and IVP SOF calves at one herd). In Study 2a, although IVP SOF calves showed (non-significant) shorter gestations, easier calvings and lower percentages of perinatal mortality and congenital malformations than IVP co-culture calves, birth weights were not decreased. In Study 2b however, the difference between IVP co-culture and IVP SOF calves in birth weight and ease of calving was significant (P < 0.05), IVP SOF calves resembling MOET calves more. IVP calves differed significantly from MOET calves with respect to several physiological parameters, such as blood oxygen saturation level, heart beat frequency and some measures of the heart. In addition, in Study 2b, recipients receiving an IVP SOF embryo showed a more regular return to estrus than those receiving an IVP co-culture embryo. From Study 2 it can be concluded that using a semi-defined medium for in vitro culture (SOF) may improve characteristics of IVP calves born.
自从牛和羊植入前胚胎的体外生产(IVP)技术问世以来,有报道称与人工授精(AI)或胚胎移植(MOET)后代相比,IVP犊牛和羔羊出生体重增加以及存在其他异常情况。本文的研究1在大规模田间条件下对AI、MOET和IVP(共培养/血清)犊牛的犊牛及产犊特征进行了比较,证实了这些报道。此外,研究表明,MOET犊牛往往出生体重更高,妊娠期明显更长,产犊也比AI犊牛更困难。目前尚不清楚IVP的影响在生命后期是否也会出现。本文展示了牛IVP共培养/血清后代的繁殖特征数据。将一岁IVP公牛的精液生产及未返情数据,以及一岁IVP母牛的超数排卵、人工授精和卵巢穿刺取卵/IVP结果,与同期生产精液或胚胎的一岁AI和MOET动物的数据进行了比较。到目前为止,没有迹象表明IVP的使用会反映在牛IVP后代异常的繁殖特征上。有人提出,在牛(和羊)胚胎的体外培养过程中使用共培养细胞和血清,可能部分解释了牛和羊IVP后代出生体重增加及其他异常情况。在半限定培养基中去除这些因素,例如合成输卵管液(SOF),可能会产生更正常的后代。研究2在田间条件(研究2a,比较AI、IVP共培养/血清和IVP SOF犊牛)和半标准化条件(研究2b,在一个牛群中比较MOET、IVP共培养/血清和IVP SOF犊牛)下对这一假设进行了研究。在研究2a中,尽管IVP SOF犊牛的妊娠期(不显著)较短,产犊更容易,围产期死亡率和先天性畸形的百分比也较低,但出生体重并未降低。然而,在研究2b中,IVP共培养犊牛和IVP SOF犊牛在出生体重和产犊难易程度上的差异显著(P < 0.05),IVP SOF犊牛更类似于MOET犊牛。IVP犊牛在一些生理参数方面与MOET犊牛有显著差异,如血氧饱和度水平、心跳频率和一些心脏指标。此外,在研究2b中,接受IVP SOF胚胎的受体母牛比接受IVP共培养胚胎的母牛发情恢复更规律。从研究2可以得出结论,使用半限定培养基(SOF)进行体外培养可能会改善出生的IVP犊牛的特征。