Embryonic stem cells (ESCs) derived from somatic cell nuclear transfer (SCNT) and induced pluripotent stem cells (iPSCs) are promising tools for meeting the personalized requirements of regenerative medicine. However, some obstacles need to be overcome before clinical trials can be undertaken. First, donor cells vary, and the reprogramming procedures are diverse, so standardization is a great obstacle regarding SCNT and iPSCs. Second, somatic cells derived from a patient may carry mitochondrial DNA mutations and exhibit telomere instability with aging or disease, and SCNT-ESCs and iPSCs retain the epigenetic memory or epigenetic modification errors. Third, reprogramming efficiency has remained low. Therefore, in addition to improving their success rate, other alternatives for producing ESCs should be explored. Producing androgenetic diploid embryos could be an outstanding strategy; androgenic diploid embryos are produced through double sperm cloning (DSC), in which two capacitated sperms (XY or XX, sorted by flow cytometer) are injected into a denucleated oocyte by intracytoplasmic sperm injection (ICSI) to reconstruct embryo and derive DSC-ESCs. This process could avoid some potential issues, such as mitochondrial interference, telomere shortening, and somatic epigenetic memory, all of which accompany somatic donor cells. Oocytes are naturally activated by sperm, which is unlike the artificial activation that occurs in SCNT. The procedure is simple and practical and can be easily standardized. In addition, DSC-ESCs can overcome ethical concerns and resolve immunological response matching with sperm providers. Certainly, some challenges must be faced regarding imprinted genes, epigenetics, X chromosome inactivation, and dosage compensation. In mice, DSC-ESCs have been produced and have shown excellent differentiation ability. Therefore, the many advantages of DSC make the study of this process worthwhile for regenerative medicine and animal breeding.

中文翻译：

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双精子克隆（DSC）是哺乳动物基因工程和干细胞研究中的一种有前途的策略。

源自体细胞核移植（SCNT）的胚胎干细胞（ESC）和诱导性多能干细胞（iPSC）是满足再生医学个性化需求的有前途的工具。但是，在进行临床试验之前需要克服一些障碍。首先，供体细胞各不相同，重新编程的程序也各不相同，因此标准化对于SCNT和iPSC来说是一个很大的障碍。其次，源自患者的体细胞可能携带线粒体DNA突变，并随着衰老或疾病表现出端粒不稳定性，并且SCNT-ESC和iPSC保留表观遗传记忆或表观遗传修饰错误。第三，重新编程效率仍然很低。因此，除了提高成功率外，还应探索生产电调的其他替代方法。产生雄激素二倍体胚胎可能是一个杰出的策略。雄激素二倍体胚胎是通过双精子克隆（DSC）产生的，其中将两个获能的精子（XY或XX，通过流式细胞仪分选）通过胞浆内精子注射（ICSI）注射到去核卵母细胞中以重建胚胎并获得DSC-ESCs。此过程可以避免一些潜在的问题，例如线粒体干扰，端粒缩短和体细胞表观遗传记忆，所有这些都伴随着体细胞供体细胞。卵母细胞被精子自然激活，这与SCNT中发生的人工激活不同。该过程简单实用，并且可以轻松标准化。另外，DSC-ESCs可以克服道德上的顾虑并解决与精子提供者匹配的免疫反应。当然，在印迹基因，表观遗传学，X染色体失活和剂量补偿方面必须面对一些挑战。在小鼠中，已经产生了DSC-ESC，并且显示出优异的分化能力。因此，DSC的许多优点使该方法的研究值得用于再生医学和动物育种。