Generation of Induced Pluripotent Stem Cells from Adult Rhesus Monkey Fibroblasts

Induced pluripotent stem (iPS) cells can be generated from somatic cells by transduction with several transcription factors in both mouse and human. However, direct reprogramming in other species has not been reported. Here, we established an efficient method to generate monkey iPS cells from fibroblasts by retrovirus-mediated introduction of the four monkey transcription factors OCT4 (POU5F1), SOX2, KLF4, and c-MYC. The monkey iPS cells displayed ES-like morphology, expressed ES cell-marker genes, shared similar global gene profiles and methylation status in the OCT4 promoter to those of monkey ES cells, and possessed the ability to differentiate into three germ layers in vitro and in vivo. Our results suggest that the mechanism of direct reprogramming is conserved among species. The efficient generation of monkey iPS cells will allow investigation of the feasibility of therapeutic cloning in primate model with various diseases. Keywords: Induced pluripotent stem, iPS, Rhesus monkey We analysed each sample (Rhesus monkey fibroblast, embryonic stem cell (ES) and induced pluripotent stem cell (iPS)) for three replications and sought to see high similarty between iPS and ES.

诱导多能干细胞（Induced pluripotent stem cells, iPS）可通过向体细胞中转导多种转录因子，在小鼠和人类中成功获得。然而目前尚未见其他物种的直接重编程相关报道。本研究通过逆转录病毒介导的方式，向恒河猴成纤维细胞中转入四种猴源转录因子OCT4（POU5F1）、SOX2、KLF4及c-MYC，建立了高效的猴诱导多能干细胞制备方法。所得猴iPS细胞呈现出与胚胎干细胞（Embryonic Stem cells, ES）相似的形态，表达ES细胞标志性基因，其全基因组表达谱与OCT4启动子区域的甲基化状态均与猴ES细胞高度相似，且具备在体外及体内分化为三个胚层的能力。本研究结果表明，直接重编程的机制在不同物种间具有保守性。高效制备猴iPS细胞的方法，将为利用罹患各类疾病的灵长类动物模型开展治疗性克隆可行性研究提供可能。关键词：诱导多能干细胞、iPS、恒河猴。本研究对每类样本（恒河猴成纤维细胞、胚胎干细胞（ES）及诱导多能干细胞（iPS））设置三次生物学重复，旨在验证iPS细胞与ES细胞之间的高度相似性。