Generation of human induced pluripotent stem cells from dermal fibroblasts
Generation of human induced pluripotent stem cells from dermal fibroblasts
复制标题
DOI：
10.1073/pnas.0711983105
复制
发表时间：
2008-02-26
影响因子：
11.1
通讯作者：
Plath, K.
中科院分区：
文献类型：
--
作者：
Lowry, W. E.;Richter, L.;Plath, K.
生成患者特异性多能干细胞有可能加速干细胞在退行性疾病临床治疗中的应用。包括体细胞核移植和细胞融合在内的技术可能产生此类细胞，但受到一些问题的阻碍，这些问题可能使其无法用于临床。在此，我们描述了利用从个体容易获取的皮肤成纤维细胞，通过异位表达特定转录因子KLF4、OCT4、SOX2和C - MYC来生成人诱导多能干细胞（iPS细胞）的方法。所得到的细胞系在形态上与人胚胎干细胞（HESC，由人植入前胚胎的内细胞团产生）无法区分。与这些观察结果一致的是，人iPS细胞与两种已建立的HESC细胞系具有几乎相同的基因表达谱。重要的是，DNA指纹图谱表明人iPS细胞来源于供体材料，并非污染所致。核型分析表明，通过特定因子对人细胞进行重编程不会诱导或需要染色体异常。最后，我们提供的证据表明，人iPS细胞可被诱导分化为代表三个胚胎胚层的细胞谱系，这表明这些细胞具有多能性。我们的研究结果是朝着操控人体体细胞以生成无限量的患者特异性多能干细胞迈出的重要一步。未来，利用特定因子改变细胞命运可能是对人体体细胞进行常规核重编程的关键。
The generation of patient-specific pluripotent stem cells has the potential to accelerate the implementation of stem cells for clinical treatment of degenerative diseases. Technologies including somatic cell nuclear transfer and cell fusion might generate such cells but are hindered by issues that might prevent them from being used clinically. Here, we describe methods to use dermal fibroblasts easily obtained from an individual human to generate human induced pluripotent stem (M) cells by ectopic expression of the defined transcription factors KLF4, OCT4, SOX2, and C-MYC. The resultant cell lines are morphologically indistinguishable from human embryonic stem cells (HESC) generated from the inner cell mass of a human preimplantation embryo. Consistent with these observations, human iPS cells share a nearly identical gene-expression profile with two established HESC lines. Importantly, DNA fingerprinting indicates that the human iPS cells were derived from the donor material and are not a result of contamination. Karyotypic analyses demonstrate that reprogramming of human cells by defined factors does not induce, or require, chromosomal abnormalities. Finally, we provide evidence that human iPS cells can be induced to differentiate along lineages representative of the three embryonic germ layers indicating the pluripotency of these cells. Our findings are an important step toward manipulating somatic human cells to generate an unlimited supply of patient-specific pluripotent stem cells. In the future, the use of defined factors to change cell fate may be the key to routine nuclear reprogramming of human somatic cells.