Sommer Cesar A, Stadtfeld Matthias, Murphy George J, Hochedlinger Konrad, Kotton Darrell N, Mostoslavsky Gustavo
Department of Medicine, Boston University School of Medicine, Massachusetts 02118, USA.
Stem Cells. 2009 Mar;27(3):543-9. doi: 10.1634/stemcells.2008-1075.
Induced pluripotent stem (iPS) cells can be generated using retroviral vectors expressing Oct4, Klf4, Sox2, and cMyc. Most prior studies have required multiple retroviral vectors for reprogramming, resulting in high numbers of genomic integrations in iPS cells and limiting their use for therapeutic applications. Here we describe the use of a single lentiviral vector expressing a "stem cell cassette" composed of the four transcription factors and a combination of 2A peptide and internal ribosome entry site technology, generating iPS cells from postnatal fibroblasts. iPS cells generated in this manner display embryonic stem cell-like morphology, express stem cell markers, and exhibit in vivo pluripotency, as evidenced by their ability to differentiate in teratoma assays and their robust contribution to mouse chimeras. Combining all factors into a single transcript achieves the most efficient reprogramming system to date and allows derivation of iPS cells with a single viral integration. The use of a single lentiviral vector for reprogramming represents a powerful laboratory tool and a significant step toward the application of iPS technology for clinical purposes.
诱导多能干细胞（iPS细胞）可以通过表达Oct4、Klf4、Sox2和cMyc的逆转录病毒载体来生成。大多数先前的研究需要多个逆转录病毒载体进行重编程，这导致iPS细胞中存在大量的基因组整合，限制了它们在治疗应用中的使用。在此，我们描述了使用一种单一的慢病毒载体，该载体表达由这四种转录因子以及2A肽和内部核糖体进入位点技术组合而成的“干细胞盒”，从出生后的成纤维细胞中生成iPS细胞。以这种方式生成的iPS细胞呈现出胚胎干细胞样的形态，表达干细胞标志物，并在体内表现出多能性，这在畸胎瘤实验中的分化能力以及对小鼠嵌合体的强大贡献中得到了证明。将所有因子整合到一个单一转录本中实现了迄今为止最有效的重编程系统，并允许通过单次病毒整合获得iPS细胞。使用单一慢病毒载体进行重编程代表了一种强大的实验室工具，也是iPS技术应用于临床目的的重要一步。
