Mouse cloning and somatic cell reprogramming using electrofused blastomeres

• Published in: Cell research Vol. 21; no. 5; pp. 770 - 778
• Main Authors: Riaz, Amjad, Zhao, Xiaoyang, Dai, Xiangpeng, Li, Wei, Liu, Lei, Wan, Haifeng, Yu, Yang, Wang, Liu, Zhou, Qi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.05.2011; Nature Publishing Group
• Subjects: 631/136/1455; 631/136/532/2435; 631/61/17/1998; Animals; Biomedical and Life Sciences; Blastomeres - cytology; Blastomeres - metabolism; Cell Biology; Cell Fusion - methods; Cell Line; Cellular Reprogramming; Chromosomes, Mammalian - metabolism; Cloning; Cloning, Organism; Cytoplasm - metabolism; Eggs; Electroporation - methods; Embryo, Mammalian - cytology; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Embryos; Fertility; Fibroblasts - cytology; Humans; Life Sciences; Mice; Mitosis; Octamer Transcription Factor-3 - metabolism; Original; original-article; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Tetraploidy; 人类胚胎干细胞; 体细胞核移植; 克隆胚胎; 卵裂球; 电熔; 胚胎干细胞系; 重编程; 鼠标; cleaved embryos; nuclear transfer; mitotic arrest; electrofusion; developmental reprogramming; MG-132
• ISSN: 1001-0602; 1748-7838
• DOI: 10.1038/cr.2010.180

Mouse cloning from fertilized eggs can assist development of approaches for the production of ＂genetically tailored＂ human embryonic stem （ES） cell lines that are not constrained by the limitations of oocyte availability. However, to date only zygotes have been successfully used as recipients of nuclei from terminally differentiated somatic cell donors leading to ES cell lines. In fertility clinics, embryos of advanced embryonic stages are usually stored for future use, but their ability to support the derivation of ES cell lines via somatic nuclear transfer has not yet been proved. Here, we report that two-cell stage electrofused mouse embryos, arrested in mitosis, can support developmental reprogramming of nuclei from donor cells ranging from blastomeres to somatic cells. Live, full-term cloned pups from embryonic donors, as well as pluripotent ES cell lines from embryonic or somatic donors, were successfully generated from these reconstructed embryos. Advanced stage pre-implantation embryos were unable to develop normally to term after electrofusion and transfer of a somatic cell nucleus, indicating that discarded pre-implantation human embryos could be an important resource for research that minimizes the ethical concerns for human therapeutic cloning. Our approach provides an attractive and practical alternative to therapeutic cloning using donated oocytes for the generation of patient-specific human ES cell lines.