Human Induced Pluripotent Stem (hiPS) Cells from Urine Samples: A Non-Integrative and Feeder-Free Reprogramming Strategy
Clara Steichen, Karim Si-Tayeb, Fanny Wulkan, Thayane Crestani, Graça Rosas, Rafael Dariolli, Alexandre C. Pereira, Jose E. Krieger
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引用次数: 16
Abstract
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.
来自尿液样本的人类诱导多能干细胞(hiPS):一种非整合和无喂食的重编程策略
人类诱导多能干细胞(hiPS)技术已经彻底改变了一些基础和应用研究,如疾病机制的研究和药理学筛选。首个使用hiPS细胞衍生细胞的临床试验开始于日本,距这篇证明概念的文章发表仅10年。在这个令人兴奋的背景下,生成hiPS细胞的策略发展迅速,趋向于非侵入性的体细胞取样方案,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,通过携带所需基因型的个体尿液样本的外泌质粒转染来产生hiPS细胞。根据先前发表的作品,这种简化的方案需要最少的设备和试剂,适合熟悉hiPS细胞技术的科学家和新手。HiPS细胞表现出多能性的经典特征,适合所有所需的下游应用,生成速度快(10周),效率高。©2017 by John Wiley &儿子,Inc。
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