Live Birth of Somatic Cell-Cloned Rabbits following Trichostatin A Treatment and Cotransfer of Parthenogenetic Embryos
Live Birth of Somatic Cell-Cloned Rabbits following Trichostatin A Treatment and Cotransfer of Parthenogenetic Embryos
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DOI：
10.1089/clo.2008.0072
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发表时间：
2009-03-01
期刊：
影响因子：
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通讯作者：
Dinnyes, Andras
中科院分区：
文献类型：
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作者：
Meng, Qinggang;Polgar, Zsuzsanna;Dinnyes, Andras
体细胞核移植(SCNT)在兔体细胞中的效率仍然较低。以往的研究表明，曲古抑素A(TSA)处理可以提高小鼠的克隆效率和长期发育，孤雌生殖(PA)胚胎的共移植有利于猪和小鼠克隆胚胎的妊娠。本研究探讨了TSA处理对SCNT兔胚胎发育的影响，以及通过共移植PA胚胎维持克隆妊娠的可能性。将卵丘细胞与去核的胞质融合，然后用电刺激、二甲氨基嘌呤(6-DMAP)和环六亚胺(CHX)处理，获得SCNT胚胎。无论是否使用TSA处理，均在EBSS-Complete培养液中培养。体外发育数据显示，TSA处理的SCNT胚胎与未处理的SCNT胚胎在卵裂率、囊胚率和囊胚细胞数上没有差异。TSA处理组的6只受体中有2只在胚胎移植(ET)后怀孕，1只怀孕的雌性动物产下了7只活幼崽和3只死胎幼崽。所有活体幼崽的死亡发生在一小时到19天内。在未接受TSA治疗的组中，七名接受者中有四人怀孕。他们中的三个生下了六只活的和八只死胎的幼崽。未经TSA处理的组中有四只幼崽已经成年，其中三只产生了后代。将3~4枚PA胚胎与26~32枚SCNT胚胎共移植给同一受体，妊娠率和出生率与对照SCNT ET组相比差异无统计学意义。综上所述，我们的结果表明，TSA处理对SCNT胚胎的体外发育影响有限；此外，TSA处理的克隆和未处理的克隆都能在兔体内发育到足月，但在我们的实验中，TSA处理的胚胎的后代都没有存活到成年。
Somatic cell nuclear transfer ( SCNT) efficiency is still low in rabbit. Previous studies indicated that trichostatin A (TSA) treatment could improve cloning efficiency and term development in the mouse, and cotransfer of parthenogenetic (PA) embryos benefited the pregnancy of cloned embryos in porcine and the mouse. In this study we investigated the effect of TSA treatment on the term development of the SCNT rabbit embryos, and the possibility of the pregnancy maintenance of clones by cotransfer of PA embryos. The SCNT embryos were produced by fusing cumulus cells with enucleated cytoplasts before activation by electrical stimulation, and Dimethylaminopurine (6-DMAP) and Cyclohexamide (CHX) treatments. They were cultured in EBSS-complete medium regardless of their treatment with or without TSA. In vitro developmental data showed no differences in the cleavage and the blastocyst rates, and the blastocyst cell number between the TSA-treated and the untreated SCNT embryos. Two of the six recipients became pregnant after the embryo transfer ( ET) in the TSA-treated group, and one pregnant female delivered seven live and three stillborn pups. The death of all live pups occurred within an hour to 19 days. Four of the seven recipients became pregnant in the TSA-untreated group. Three of them gave birth to six live and eight stillborn pups. Four pups of the TSA-untreated group have grown into adulthood, and three of them produced progeny. Cotransfer of three to four PA embryos with 26-32 SCNT embryos to the same recipient resulted in pregnancy and birth rates statistically no different compared to the control SCNT ET group. In conclusion, our results indicate that TSA treatment has a limited effect on the in vitro development of the SCNT embryos; furthermore, both the TSA-treated and the untreated clones can develop to term in rabbits, but none of the offspring from TSA-treated embryos survived to adulthood in our experiment.