iPS-technology and patient specific disease models

IPS技术的发展和应用有望彻底改变对疾病的理解和治疗，因为重新编程患者特定的iPS细胞的能力使基于细胞的疾病模型的建立成为可能，在这种疾病模型中，以前很难获得足够数量的相关人类细胞类型，包括心肌细胞、内皮细胞和血管平滑肌细胞，以探索心血管疾病的潜在机制。我们已经成功地从健康志愿者和患者身上培养出了几个人类iPS细胞系。到目前为止，这些患者特有的细胞系包括乔布斯综合征、CADASIL、DADA2、特纳综合征和Degos患者，仅举几例。我们已经成功地使用慢病毒和逆转录病毒传递四种必需的多能因子，并广泛地表征了许多产生的人类iPS细胞系。我们的iPS细胞系表现出人类胚胎干细胞(ESC)的特征，包括形成刺状体的能力。这些细胞还表达高水平的碱性磷酸酶、Oct4和Nanog，以及人类ESC标记SSEA4和Tra-1-60。此外，我们已经证实这些细胞表现出正常的核型，并制定了将这些iPS细胞向不同的外胚层、中胚层和内胚层分化的方案，通过Nestin、RUNX1和GATA4的表达来评估。IPS细胞已经成功地分化为与心血管疾病相关的细胞系，包括内皮细胞和平滑肌细胞，我们正在继续将它们应用于我们研究的疾病的体外模型中。

The development and application of iPS technology promises to revolutionize the understanding and treatment of disease, as the ability to reprogram patient specific iPS cells permit the creation of cell-based disease models where it has previously been extremely difficult to obtain sufficient amounts of the relevant human cell types, including cardiac myocytes, endothelial cells and vascular smooth muscle cells, to explore the mechanisms underlying cardiovascular diseases. We have successfully generated several human iPS cell lines from both healthy volunteers and patients. To date, these patient specific cell lines have included individuals with Jobs syndrome, CADASIL, DADA2, Turner Syndrome and Degos just to name a few. We have been successful using both lentiviral and retroviral delivery of the four requisite pluripotent factors and have also extensively characterized many of the human iPS cell lines generated. Our iPS cell lines exhibit characteristics of human embryonic stem cells (ESC) including the ability to form embroid bodies. These cells also express high levels of alkaline phosphatase, Oct4 and Nanog, as well as the human ESC markers SSEA4 and Tra-1-60. Furthermore, we have confirmed that these cells manifest a normal karyotype and have developed protocols for differentiation of these iPS cells down various ectodermal, mesodermal and endodermal lineages as assessed by the respective expression of Nestin, Runx1 and GATA4. Successful differentiation of iPS cells towards lineages that are relevant for cardiovascular disease, including endothelial and smooth muscle cells, has been achieved and we are continuing to apply them in in vitro models of diseases that we investigate.