An intracytoplasmic injection of deionized bovine serum albumin immediately after somatic cell nuclear transfer enhances full-term development of cloned mouse embryos.
An intracytoplasmic injection of deionized bovine serum albumin immediately after somatic cell nuclear transfer enhances full-term development of cloned mouse embryos.
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DOI：
10.1262/jrd.2015-031
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Yamada M
中科院分区：
文献类型：
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作者：
Isaji Y;Yoshida K;Imai H;Yamada M
在小鼠体细胞核移植（SCNT）中，聚乙烯吡咯烷酮（PVP）通常包含在核供体注射介质中。然而，将PVP注射到卵母细胞的细胞质中，其细胞毒性最近引起了人们的关注。在本研究中，我们确定了用离子交换树脂处理（d-BSA）去离子牛血清白蛋白是否适用于SCNT的核供体注射介质，作为PVP的替代品。结果表明，将d-BSA与供体核一起注入去核卵母细胞的细胞质中，与常规核注射PVP相比，可显著提高克隆胚胎在体外发育到囊胚期的速度。在卵母细胞激活前，将d-BSA注射到与日本包膜血凝病毒融合法重建的卵母细胞的细胞质中，观察其对囊胚形成率的增强作用。免疫荧光实验显示，注射d-BSA可提高克隆原核（PN）和2细胞胚胎中组蛋白H3赖氨酸9和H4赖氨酸12的乙酰化水平。在卵母细胞激活前注射d-BSA也增加了克隆小鼠后代的产量。上述结果提示，在卵母细胞活化前，胞浆内注射d-BSA有利于通过表观遗传修饰核重编程促进小鼠克隆胚胎的体外和体内发育。
In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in vitro development of cloned embryos to the blastocyst stage compared with that of a conventional nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine 12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear reprogramming.