title: Mouse cloning and somatic cell reprogramming using electrofused blastomeres.
abstract: 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.
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 1st Beichen West Road, Beijing 100101, China.
authors.forename: Amjad
authors.initials: A
authors.lastname: Riaz
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiangpeng
authors.initials: X
authors.lastname: Dai
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.forename: Haifeng
authors.initials: H
authors.lastname: Wan
authors.forename: Yang
authors.initials: Y
authors.lastname: Yu
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001757:Blastomeres / Q000166:cytology* / Q000378:metabolism; D002459:Cell Fusion / Q000379:methods*; D002460:Cell Line; D065150:Cellular Reprogramming; D033481:Chromosomes, Mammalian / Q000378:metabolism; D019976:Cloning, Organism; D003593:Cytoplasm / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D051379:Mice; D008938:Mitosis; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D057891:Tetraploidy
chemical_list: D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse