SUCCESSFUL IN-VITRO CULTURE OF EARLY CLEAVAGE STAGE EMBRYOS RECOVERED FROM SUPEROVULATED RED DEER (CERVUS-ELAPHUS)
SUCCESSFUL IN-VITRO CULTURE OF EARLY CLEAVAGE STAGE EMBRYOS RECOVERED FROM SUPEROVULATED RED DEER (CERVUS-ELAPHUS)
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DOI：
10.1016/0093-691x(95)00174-7
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发表时间：
1995-07-15
期刊：
影响因子：
2.8
通讯作者：
ASHER, GW
中科院分区：
文献类型：
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作者：
BERG, DK;THOMPSON, JG;ASHER, GW
评价了三种单独的胚胎培养系统支持早期卵裂期红鹿（马鹿）胚胎发育的能力：结扎的绵羊输卵管（处理A）; TCM 199 + 10%鹿血清中的鹿输卵管上皮单层（处理B）; 7%O-2气氛下的合成输卵管液+ 20%人血清（处理C）。此外，还比较了两种超数排卵方案在产生早期卵裂期胚胎方面的效果。20头红鹿（2 ~ 7岁）在4月用阴道内CIDR（TM）装置同步化12天。所有动物在取出CIDR（TM）装置前72小时开始接受8次相等剂量的0.4单位绵羊FSH给药，间隔12小时。鹿另外接受200 IU PMSG，与第一次FSH注射（组1，n=10）或与最后一次FSH注射（组2，n=10）。在取出CIDR装置后，将母鹿与可育雄鹿一起放置。在CIDR去除后63小时通过手术回收收集卵。在收集时，组2中的动物具有显著更高的平均（+/- SEM）排卵率（11.2+/-2.4 vs 5.3 + 2.4），其中比组1中的动物（10/10 vs 4/10）更多的动物响应于处理（>1次排卵）。在繁殖季节后期（6月），使用与组2中鹿相同的方案对另外10只红鹿（组3，实验2）进行超数排卵，提前24 h收集卵子。平均（+/- SEM）排卵率为6.4 +/- 1.2，9/10只动物有反应。各组之间的卵回收率没有差异（范围为73 - 87%）。超数排卵处理不影响培养胚胎发育到桑椹胚/囊胚期。此外，3种培养系统对桑椹胚（范围50 - 58%）或胚泡（范围22 - 26%）阶段的支持率无差异。在腹腔镜下将4个桑椹胚/囊胚胚胎移植至受体红鹿（2个来自处理组B，2个来自处理组C）后，从处理组B中培养的胚胎产下2头活小牛。
Three separate embryo culture systems were evaluated for their ability to support development of early cleavage stage red deer (Cervus elaphus) embryos: ligated sheep oviducts (Treatment A); cervine oviduct epithelial monolayer in TCM 199 + 10% deer serum (Treatment B); synthetic oviduct fluid + 20% human serum at 7% O-2 atmosphere (Treatment C). In addition, 2 superovulation protocols were compared for their efficacy in producing early cleavage stage embryos. Twenty red deer (2 to 7 yr old) were synchronized in April with intravaginal CIDR(TM) devices for 12 d. All animals received a total of 0.4 units of ovine FSH administered in 8 equal doses, 12 h apart, beginning 72 h before removal of CIDR(TM) devices. The deer additionally received 200 IU PMSG, either with the first FSH injection (Group 1, n=10) or with the last FSH injection (Group 2, n=10). Hinds were placed with fertile stags following withdrawal of CIDR devices. Ova were collected by surgical recovery 63 h post CIDR removal. At the time of collection, animals in Group 2 had a significantly greater mean (+/- SEM) ovulation rate (11.2+/-2.4 vs 5.3 + 2.4), with more animals responding to treatment (>1 ovulation), than the animals in Group 1 (10/10 vs 4/10). Late in the breeding season(June), 10 additional red deer (Group 3, Experiment 2) were superovulated using the same protocol as for the deer in Group 2, with ova collection advanced by 24 h. Mean (+/- SEM) ovulation rate was 6.4 +/- 1.2 with 9/10 animals responding. Ova recovery did not differ among the groups (range 73 to 87%). Superovulation treatment did not affect cultured embryo development to the morula/blastocyst stage. Furthermore, there was no difference among the 3 culture systems in their support of development either to the morula (range 50 to 58%) or to the blastocyst (range 22 to 26%) stage. After laparoscopic transfer of 4 morula/blastocyst embryos to recipient red deer (2 from Treatment B and 2 from Treatment C) 2 live calves were born from embryos cultured in Treatment B.