Steichen Clara, Si-Tayeb Karim, Wulkan Fanny, Crestani Thayane, Rosas Graça, Dariolli Rafael, Pereira Alexandre C, Krieger Jose E
Heart Institute (InCor), University of São Paulo Medical School, São Paulo, Brazil.
INSERM, UMR1087, L'Institut du Thorax, Nantes, France.
Curr Protoc Hum Genet. 2017 Jan 11;92:21.7.1-21.7.22. doi: 10.1002/cphg.26.
Human induced pluripotent stem (hiPS) cell technology has already revolutionized some aspects of fundamental and applied research such as study of disease mechanisms and pharmacology screening. The first clinical trial using hiPS cell-derived cells began in Japan, only 10 years after the publication of the proof-of concept article. In this exciting context, strategies to generate hiPS cells have evolved quickly, tending towards non-invasive protocols to sample somatic cells combined with "safer" reprogramming strategies. In this unit, we describe a protocol combining both of these advantages to generate hiPS cells with episomal plasmid transfection from urine samples of individuals carrying the desired genotype. Based on previous published works, this simplified protocol requires minimal equipment and reagents, and is suitable both for scientists familiar with the hiPS cells technology and neophytes. HiPS cells displaying classical features of pluripotency and suitable for all desired downstream applications are generated rapidly (<10 weeks) and with high efficiency. © 2017 by John Wiley & Sons, Inc.
人类诱导多能干细胞（hiPS）技术已经在基础研究和应用研究的某些方面引发了变革，比如疾病机制研究和药理学筛选。在概念验证文章发表仅10年后，日本就开展了首例使用hiPS细胞衍生细胞的临床试验。在这种令人兴奋的背景下，生成hiPS细胞的策略迅速发展，倾向于采用非侵入性方案来获取体细胞，并结合“更安全”的重编程策略。在本单元中，我们描述了一种结合这两种优势的方案，即通过携带所需基因型个体的尿液样本，利用游离质粒转染来生成hiPS细胞。基于先前发表的研究成果，这种简化方案所需的设备和试剂最少，既适合熟悉hiPS细胞技术的科学家，也适合新手。能快速（<10周）且高效地生成具有多能性经典特征且适用于所有所需下游应用的hiPS细胞。© 2017约翰威立国际出版公司
