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Human embryonic stem cell lines derived from single blastomeres

机译：源自单个卵裂球的人类胚胎干细胞系

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The derivation of human embryonic stem (hES) cells currently requires the destruction of ex utero embryos(1-4). A previous study in mice indicates that it might be possible to generate embryonic stem (ES) cells using a single-cell biopsy similar to that used in preimplantation genetic diagnosis (PGD), which does not interfere with the embryo's developmental potential(5). By growing the single blastomere overnight, the resulting cells could be used for both genetic testing and stem cell derivation without affecting the clinical outcome of the procedure. Here we report a series of ten separate experiments demonstrating that hES cells can be derived from single blastomeres. In this proof-of-principle study, multiple biopsies were taken from each embryo using micromanipulation techniques and none of the biopsied embryos were allowed to develop in culture. Nineteen ES-cell-like outgrowths and two stable hES cell lines were obtained. The latter hES cell lines maintained undifferentiated proliferation for more than eight months, and showed normal karyotype and expression of markers of pluri-potency, including Oct-4, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, nanog and alkaline phosphatase. These cells retained the potential to form derivatives of all three embryonic germ layers both in vitro and in teratomas. The ability to create new stem cell lines and therapies without destroying embryos would address the ethical concerns of many, and allow the generation of matched tissue for children and siblings born from transferred PGD embryos.
机译：目前人类胚胎干（hES）细胞的衍生需要破坏子宫外胚胎（1-4）。一项先前在小鼠中的研究表明，与植入前遗传学诊断（PGD）中使用的类似，使用单细胞活检可能会产生胚胎干（ES）细胞，这不会干扰胚胎的发育潜力（5）。通过使单个卵裂球生长过夜，可以将所得细胞用于基因测试和干细胞衍生，而不会影响该过程的临床结果。在这里，我们报告一系列的十个独立的实验，证明hES细胞可以来自单个卵裂球。在这项原理验证研究中，使用显微操作技术从每个胚胎中进行了多次活检，没有任何活检胚胎在培养物中发育。获得了十九个类似ES细胞的产物和两个稳定的hES细胞系。后者的hES细胞系保持未分化的增殖超过八个月，并显示出正常的核型和多潜能标志物的表达，包括Oct-4，SSEA-3，SSEA-4，TRA-1-60，TRA-1- 81，纳微和碱性磷酸酶。这些细胞保留了在体外和畸胎瘤中形成所有三个胚芽层的衍生物的潜力。在不破坏胚胎的情况下创建新的干细胞系和疗法的能力将解决许多伦理问题，并允许为从转移的PGD胚胎出生的孩子和兄弟姐妹生成匹配的组织。

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