Wakayama Sayaka, Jakt Martin L, Suzuki Masako, Araki Ryoko, Hikichi Takafusa, Kishigami Satoshi, Ohta Hiroshi, Van Thuan Nguyen, Mizutani Eiji, Sakaide Yuko, Senda Sho, Tanaka Satoshi, Okada Mitsuhiro, Miyake Masashi, Abe Masumi, Nishikawa Shin-Ichi, Shiota Kunio, Wakayama Teruhiko
Laboratory for Genomic Programming, RIKEN Center for Developmental Biology, Kobe, Japan.
Stem Cells. 2006 Sep;24(9):2023-33. doi: 10.1634/stemcells.2005-0537. Epub 2006 May 11.
Therapeutic cloning, whereby nuclear transfer (NT) is used to generate embryonic stem cells (ESCs) from blastocysts, has been demonstrated successfully in mice and cattle. However, if NT-ESCs have abnormalities, such as those associated with the offspring produced by reproductive cloning, their scientific and medical utilities might prove limited. To evaluate the characteristics of NT-ESCs, we established more than 150 NT-ESC lines from adult somatic cells of several mouse strains. Here, we show that these NT-ESCs were able to differentiate into all functional embryonic tissues in vivo. Moreover, they were identical to blastocyst-derived ESCs in terms of their expression of pluripotency markers in the presence of tissue-dependent differentially DNA methylated regions, in DNA microarray profiles, and in high-coverage gene expression profiling. Importantly, the NT procedure did not cause irreversible damage to the nuclei. These similarities of NT-ESCs and ESCs indicate that murine therapeutic cloning by somatic cell NT can provide a reliable model for preclinical stem cell research.
治疗性克隆是指通过核移植（NT）从囊胚中生成胚胎干细胞（ESC），这已在小鼠和牛身上成功得到证实。然而，如果NT-ESC存在异常，比如与生殖克隆所产生的后代相关的那些异常，那么它们在科学和医学上的用途可能会很有限。为了评估NT-ESC的特性，我们从几种小鼠品系的成年体细胞中建立了150多个NT-ESC系。在此，我们表明这些NT-ESC能够在体内分化为所有功能性胚胎组织。此外，在存在组织依赖性差异DNA甲基化区域的情况下、在DNA微阵列图谱中以及在高覆盖度基因表达谱中，它们在多能性标志物的表达方面与囊胚来源的ESC相同。重要的是，NT程序不会对细胞核造成不可逆的损伤。NT-ESC和ESC的这些相似性表明，通过体细胞NT进行的小鼠治疗性克隆可为临床前干细胞研究提供一个可靠的模型。
