Differentiation of Human Induced Pluripotent Stem Cells into Keratinocytes.
Differentiation of Human Induced Pluripotent Stem Cells into Keratinocytes.
复制标题
DOI：
10.1002/cpz1.408
复制
发表时间：
2022-04
期刊：
影响因子：
--
通讯作者：
Koster, Maranke I
中科院分区：
文献类型：
--
作者：
Koch, Peter J;Webb, Saiphone;Gugger, Jessica A;Salois, Maddison N;Koster, Maranke I
研究人类疾病的基本生物学机制依赖于足够数量的患者细胞的可用性。由于大多数原代体细胞的寿命有限，因此获得足够的生物学研究材料一直是一个挑战。诱导多能干细胞（iPSC）技术的发展已经改变了游戏规则，特别是在罕见遗传疾病领域。iPSC基本上是永生的，可以无限期地储存，因此可以用于无限量地产生限定的体细胞。此外，基因组编辑技术的可用性，如CRISPR/CAS，为我们提供了创造具有明确遗传特征的“设计师”iPSC系的机会。生物学研究的一个重大进展源于指导iPSC分化为定义的细胞类型的方法的发展。在这篇文章中，我们提供了产生人iPSC衍生的角质形成细胞（iPSC-K）的基本方案。这些细胞具有基底表皮角质形成细胞的特征，代表了研究正常表皮生物学以及遗传性和获得性皮肤病的工具。版权所有© 2022作者。由Wiley Periodicals LLC出版。 基本方案：支持方案1：用玻连蛋白XF™包被细胞培养皿或板 支持方案2：冷冻iPSC支持方案3：制备AggreWell™400 6孔板用于EB形成支持方案4：用胶原IV包被细胞培养皿或板支持方案5：细胞的免疫荧光染色
Investigating basic biological mechanisms underlying human diseases relies on the availability of sufficient quantities of patient cells. As most primary somatic cells have a limited lifespan, obtaining sufficient material for biological studies has been a challenge. The development of induced pluripotent stem cell (iPSC) technology has been a game changer, especially in the field of rare genetic disorders. iPSC are essentially immortal, can be stored indefinitely, and can thus be used to generate defined somatic cells in unlimited quantities. Further, the availability of genome editing technologies, such as CRISPR/CAS, has provided us with the opportunity to create “designer” iPSC lines with defined genetic characteristics. A major advancement in biological research stems from the development of methods to direct iPSC differentiation into defined cell types. In this article, we provide the basic protocol for the generation of human iPSC‐derived keratinocytes (iPSC‐K). These cells have the characteristics of basal epidermal keratinocytes and represent a tool for the investigation of normal epidermal biology, as well as genetic and acquired skin disorders. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Directed differentiation of human iPSC into keratinocytes Support Protocol 1: Coating cell culture dishes or plates with Vitronectin XF™ Support Protocol 2: Freezing iPSC Support Protocol 3: Preparing AggreWell™400 6‐well plates for EB formation Support Protocol 4: Coating cell culture dishes or plates with Collagen IV Support Protocol 5: Immunofluorescence staining of cells