Generation of germline-competent induced pluripotent stem cells

已有 365 次阅读 2010-4-13 08:16 |个人分类:literature express|关键词:stem Generation cells induced pluripotent

Nature 2007.

Abstract:

We have previously shown that pluripotent stem cells can be induced from mouse fibroblasts by retroviral introduction of Ocr3/4 (all called Pou5f1), Sox2, c-Myc and Klf4, and subsequent selection for Fbx15 (also called Fbxo15) expression. These induced pluripotent stem (iPS) cells (hereafter called Fbx15 iPS cells) are similar to embryonic stem (ES) cells in morphology, proliferation, and teratoma formation; however, they are different with regards to gene expression and DNA methylation paterns, and fail to produce adult chimaeras. Here we show that selection for Nanog expression results in germline-competent iPS cells with increased ES-cell-like gene expression and DNA methylation patterns compared with Fbx15 iPS cells. The four transgene (Oct3/4, Sox2. c-Myc and Klf4) were strongly silenced in Nanog iPS cell. We obtained adult chimaeras from seven Nanog iPS cell clones, with one clone being transmitted through the germ line to the next generation. Approximately 20% of the offspring developed tumours attributable to reactivation of the c-myc transgene. Thus, iPS cells competent for germline chimaeras can be obtained from fibroblasts, but reroviral introduction of c-Myc should be avoid for clinical application.