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灵长类动物中通过体细胞核移植进行的表观遗传重编程。

Epigenetic reprogramming by somatic cell nuclear transfer in primates.

作者信息

Sparman Michelle, Dighe Vikas, Sritanaudomchai Hathaitip, Ma Hong, Ramsey Cathy, Pedersen Darlene, Clepper Lisa, Nighot Prashant, Wolf Don, Hennebold Jon, Mitalipov Shoukhrat

机构信息

Division of Reproductive Sciences, Oregon National Primate Research Center, School of Medicine, Oregon Health and Science University, Beaverton, Oregon, USA.

出版信息

Stem Cells. 2009 Jun;27(6):1255-64. doi: 10.1002/stem.60.

Abstract

We recently demonstrated that somatic cells from adult primates could be reprogrammed into a pluripotent state by somatic cell nuclear transfer. However, the low efficiency with donor cells from one monkey necessitated the need for large oocyte numbers. Here, we demonstrate nearly threefold higher blastocyst development and embryonic stem (ES) cell derivation rates with different nuclear donor cells. Two ES cell lines were isolated using adult female rhesus macaque skin fibroblasts as nuclear donors and oocytes retrieved from one female, following a single controlled ovarian stimulation. In addition to routine pluripotency tests involving in vitro and in vivo differentiation into various somatic cell types, primate ES cells derived from reprogrammed somatic cells were also capable of contributing to cells expressing markers of germ cells. Moreover, imprinted gene expression, methylation, telomere length, and X-inactivation analyses were consistent with accurate and extensive epigenetic reprogramming of somatic cells by oocyte-specific factors.

摘要

我们最近证明,成年灵长类动物的体细胞可通过体细胞核移植重编程为多能状态。然而,来自一只猴子的供体细胞效率较低,需要大量的卵母细胞。在此,我们证明使用不同的核供体细胞,囊胚发育率和胚胎干细胞(ES细胞)衍生率提高了近三倍。使用成年雌性恒河猴皮肤成纤维细胞作为核供体,并在单次控制性卵巢刺激后从一只雌性体内获取卵母细胞,分离出了两条ES细胞系。除了涉及体外和体内分化为各种体细胞类型的常规多能性测试外,源自重编程体细胞的灵长类ES细胞也能够分化为表达生殖细胞标志物的细胞。此外,印记基因表达、甲基化、端粒长度和X染色体失活分析与卵母细胞特异性因子对体细胞进行准确而广泛的表观遗传重编程相一致。

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