Lee Sang-Goo, Park Jin-Kyu, Choi Kwang-Hwan, Son Hye-Young, Lee Chang-Kyu
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-921, Korea ; Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151-921, Korea ; Division of Animal Sciences and Bond Life Sciences Center, University of Missouri, Columbia, MO 65211, USA.
Asian-Australas J Anim Sci. 2015 Nov;28(11):1565-72. doi: 10.5713/ajas.15.0462.
Porcine embryonic stem cells (pESCs) have become an advantageous experimental tool for developing therapeutic applications and producing transgenic animals. However, despite numerous reports of putative pESC lines, deriving validated pESC lines from embryos produced in vitro remains difficult. Here, we report that embryo aggregation was useful for deriving pESCs from in vitro-produced embryos. Blastocysts derived from embryo aggregation formed a larger number of colonies and maintained cell culture stability. Our derived cell lines demonstrated expression of pluripotent markers (alkaline phosphatase, Oct4, Sox2, and Nanog), an ability to form embryoid bodies, and the capacity to differentiate into the three germ layers. A cytogenetic analysis of these cells revealed that all lines derived from aggregated blastocysts had normal female and male karyotypes. These results demonstrate that embryo aggregation could be a useful technique to improve the efficiency of deriving ESCs from in vitro-fertilized pig embryos, studying early development, and deriving pluripotent ESCs in vitro in other mammals.
猪胚胎干细胞（pESCs）已成为开发治疗应用和生产转基因动物的有利实验工具。然而，尽管有许多关于假定pESC系的报道，但从体外产生的胚胎中获得经过验证的pESC系仍然很困难。在此，我们报告胚胎聚集对于从体外产生的胚胎中获得pESCs很有用。胚胎聚集产生的囊胚形成了更多的集落并维持了细胞培养稳定性。我们获得的细胞系显示出多能性标志物（碱性磷酸酶、Oct4、Sox2和Nanog）的表达、形成胚状体的能力以及分化为三个胚层的能力。对这些细胞的细胞遗传学分析表明，所有来自聚集囊胚的细胞系都具有正常的雌性和雄性核型。这些结果表明，胚胎聚集可能是一种有用的技术，可提高从体外受精猪胚胎中获得ESCs的效率、研究早期发育以及在体外从其他哺乳动物中获得多能ESCs的效率。
