Thompson J G, Gardner D K, Pugh P A, McMillan W H, Tervit H R
AgResearch, Ruakura Agricultural Centre, Hamilton, New Zealand.
Biol Reprod. 1995 Dec;53(6):1385-91. doi: 10.1095/biolreprod53.6.1385.
It has previously been reported that ovine embryos cultured in Synthetic Oviduct Fluid medium supplemented with 20% human serum (SOF+HS) develop into lambs with a high birth weight. We have investigated this phenomenon by culturing ovine zygotes in SOF+HS or a serum-free version of Synthetic Oviduct Fluid with BSA and amino acids (SOFaaBSA) in place of serum. Zygotes were either obtained from superovulated and naturally mated ewes or produced in vitro. Embryos were subsequently transferred to synchronized recipient ewes (n = 63). An additional group of ewes (n = 16) served as flock fertility and lambing controls. Development of zygotes to stages suitable for transfer (i.e., good to excellent compact morulae or blastocysts) was not affected by medium (SOFaaBSA = 53 +/- 5% vs. SOF+HS = 59 +/- 5%) but was affected by source (in vivo-derived = 74 +/- 5% vs. in vitro-derived = 35 +/- 5%, p < 0.001). Embryos incubated in SOF+HS were morphologically different from those incubated in SOFaaBSA, having abundant lipid droplets. Pregnancy rate (65%) and embryo survival (48%) of recipients determined by ultrasonography on approximately Day 60 of pregnancy did not differ between medium treatments or source of embryo. Mean weight of lambs from embryos cultured in SOF+HS (4.2 +/- 0.2 kg) was significantly heavier than that of controls (3.4 +/- 0.2 kg, p < 0.01) or of lambs from embryos cultured in SOFaaBSA (3.5 +/- 0.2 kg, p < 0.05). Furthermore, mean gestation length was longer in recipients receiving embryos incubated in SOF+HS (147 +/- 1 days) than in SOFaaBSA (145 +/- 1 day, p < 0.05). Reasons for this birth weight and gestation length difference are unclear, but our data suggest that different culture conditions can produce embryos with differing morphology, apparent chemical composition, and rate of development, resulting in lambs with differing gestation length and birth weight.
此前有报道称,在添加20%人血清的合成输卵管液培养基(SOF+HS)中培养的绵羊胚胎发育成的羔羊出生体重较高。我们通过在SOF+HS或不含血清但添加了牛血清白蛋白和氨基酸的合成输卵管液(SOFaaBSA)中培养绵羊受精卵来研究这一现象。受精卵要么从经超排处理并自然交配的母羊获取,要么体外产生。随后将胚胎移植到同期发情的受体母羊(n = 63)体内。另外一组母羊(n = 16)作为羊群繁殖力和产羔对照。受精卵发育到适合移植阶段(即良好至优秀的致密桑葚胚或囊胚)不受培养基影响(SOFaaBSA = 53±5% 对比 SOF+HS = 59±5%),但受来源影响(体内来源 = 74±5% 对比 体外来源 = 35±5%,p < 0.001)。在SOF+HS中培养的胚胎与在SOFaaBSA中培养的胚胎形态不同,前者有丰富的脂滴。在妊娠约第60天时通过超声检查确定的受体母羊妊娠率(65%)和胚胎存活率(48%)在不同培养基处理或胚胎来源之间没有差异。在SOF+HS中培养的胚胎所产羔羊的平均体重(4.2±0.2千克)显著高于对照组(3.4±0.2千克,p < 0.01)或在SOFaaBSA中培养的胚胎所产羔羊(3.5±0.2千克,p < 0.05)。此外,接受在SOF+HS中培养的胚胎的受体母羊平均妊娠期(147±1天)比接受在SOFaaBSA中培养的胚胎的受体母羊长(145±1天,p < 0.05)。这种出生体重和妊娠期差异的原因尚不清楚,但我们的数据表明,不同的培养条件会产生形态、表观化学成分和发育速率不同的胚胎,从而导致妊娠期和出生体重不同的羔羊。