Comparison of Reprogramming Efficiency Between Transduction of Reprogramming Factors, Cell-Cell Fusion, and Cytoplast Fusion
Comparison of Reprogramming Efficiency Between Transduction of Reprogramming Factors, Cell-Cell Fusion, and Cytoplast Fusion
复制标题
DOI：
10.1002/stem.466
复制
发表时间：
2010-08-01
期刊：
影响因子：
5.2
通讯作者：
Pera, Martin F.
中科院分区：
文献类型：
--
作者：
Hasegawa, Kouichi;Zhang, Peilin;Pera, Martin F.
将人类体细胞重新编程为多能细胞为移植治疗、疾病研究和药物筛选开辟了新的可能性。除了体细胞核移植，已经报道了几种对人类细胞重新编程的方法：转导特定的转录因子产生诱导多能干细胞(IPSC)，人胚胎干细胞(HESC)体细胞融合，hESC胞质-体细胞融合或暴露于hESC提取物。在这里，我们优化了hESC-人成纤维细胞融合、去核和胞质融合的技术，并比较了IPSC生成、细胞融合和胞质融合的重编程效率。与IPSC相比，hESC-Fusion提供了更快和更有效的体细胞重编程。重编程需要4周以上，IPSC生成的效率低于0.001%，hESC融合的效率小于10天，大于0.005%。此外，融合几乎没有产生部分重新编程的细胞克隆。融合细胞为四倍体或非整倍体。HESC胞质融合可启动重编程，但始终不能完成重编程。这些数据表明，在细胞融合中，就像在核移植中一样，通过将体细胞核直接引入多能细胞的环境中进行重编程提供了相对有效的重编程。这些发现还表明，宿主多能细胞的细胞核可能含有加速重新编程过程的成分。干细胞2010；28：1338-1348
Reprogramming human somatic cells into pluripotent cells opens up new possibilities for transplantation therapy, the study of disease, and drug screening. In addition to somatic cell nuclear transfer, several approaches to reprogramming human cells have been reported: transduction of defined transcription factors to generate induced pluripotent stem cell (iPSC), human embryonic stem cell (hESC)somatic cell fusion, and hESC cytoplast-somatic cell fusion or exposure to extracts of hESC. Here, we optimized techniques for hESC-human fibroblast fusion and enucleation and cytoplast fusion, and then compared the reprogramming efficiency between iPSC generation, cell-fusion and cytoplast-fusion. When compared with iPSC, hESC-fusion provided much faster and efficient reprogramming of somatic cells. The reprogramming required more than 4 weeks and the efficiency was less than 0.001% in iPSC generation, and it was less than 10 days and more than 0.005% in hESC-fusion. In addition, fusion yielded almost no partially reprogrammed cell colonies. However, the fused cells were tetraploid or aneuploid. hESC cytoplast fusion could initiate reprogramming but was never able to complete reprogramming. These data indicate that in cell fusion, as in nuclear transfer, reprogramming through direct introduction of a somatic nucleus into the environment of a pluripotent cell provides relatively efficient reprogramming. The findings also suggest that the nucleus of the host pluripotent cell may contain components that accelerate the reprogramming process. STEM CELLS 2010; 28: 1338-1348