Kuwayama Hiroki, Tanabe Yoshiaki, Wakayama Teruhiko, Kishigami Satoshi
Faculty of Life and Environmental Sciences, University of Yamanashi, Yamanashi, Japan.
Faculty of Life and Environmental Sciences, University of Yamanashi, Yamanashi, Japan; Advanced Biotechnology Center, University of Yamanashi, 4-4-37, Takeda, Kofu-shi, Yamanashi, Japan.
Theriogenology. 2017 May;94:79-85. doi: 10.1016/j.theriogenology.2017.02.012. Epub 2017 Feb 20.
Less invasive methods for donor cell collection will facilitate reproduction of wild animals using somatic-cell nuclear transfer. Stages of the estrous cycle in mice have long been studies using somatic cells that can be collected from vaginal walls using cotton tipped swabs in a relatively non-invasive manner. In this study, we examined the feasibility of these cells as sources of nuclei for somatic-cell cloning using nuclear transfer. Estrous cycles generally comprise proestrus, estrus, metestrus, and diestrus stages. In the present experiments, more than 60% of cells were nucleated in vaginal smears from all but the estrus stage. However, after somatic-cell nuclear transfer of cells from proestrus, metestrus, and diestrus stages, 66%, 50%, and 72% of cloned embryos developed to the morula/blastocyst, and cloned female mouse birth rates after embryo transfer were 1.5%, 0.3%, and 1%, respectively. These results show that noninvasively collected vaginal smears contain somatic cells that can be used to clone female mice.
用于供体细胞采集的侵入性较小的方法将有助于利用体细胞核移植技术繁殖野生动物。长期以来,人们一直利用体细胞研究小鼠发情周期的各个阶段,这些体细胞可以通过棉拭子以相对非侵入性的方式从阴道壁采集。在本研究中,我们研究了这些细胞作为体细胞核移植克隆细胞核来源的可行性。发情周期通常包括发情前期、发情期、发情后期和间情期。在本实验中,除发情期外,所有阴道涂片中有超过60%的细胞具有细胞核。然而,在对发情前期、发情后期和间情期的细胞进行体细胞核移植后,分别有66%、50%和72%的克隆胚胎发育到桑椹胚/囊胚阶段,胚胎移植后克隆雌性小鼠的出生率分别为1.5%、0.3%和1%。这些结果表明,通过非侵入性采集的阴道涂片含有可用于克隆雌性小鼠的体细胞。