Incyte Research Institute, Wilmington, Delaware.
DermTech, La Jolla, California.
Curr Protoc. 2021 Mar;1(3):e72. doi: 10.1002/cpz1.72.
Skin is made up of an epidermis and, dermis which serve as a barrier against physical and environmental threats. Keratinocytes make up greater than 95% of the epidermis and form different layers based on their level of differentiation. Millions of individuals suffer from skin diseases, which are characterized by significant barrier disruption and inflammation. Investigators previously relied on animal models to investigate inflammatory skin diseases; however, technological advances have enabled the use of physiologically human skin models to investigate the effects of inflammatory mediators on the structure and function of skin cells. In this article, we describe two protocols using keratinocytes to investigate tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) driven skin inflammation as a surrogate for psoriasis, vitiligo, and other autoimmune skin diseases driven by these cytokines. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Preparing a HaCaT keratinocyte culture Basic Protocol 2: 3-Dimensional organotypic skin cultures to assess TNF-α and IFN-γ driven skin inflammation.
皮肤由表皮和真皮组成,它们作为抵御物理和环境威胁的屏障。角质形成细胞构成表皮的 95%以上,并根据其分化程度形成不同的层。数以百万计的人患有皮肤疾病,其特征是明显的屏障破坏和炎症。研究人员以前依赖动物模型来研究炎症性皮肤疾病;然而,技术进步使得能够使用生理上的人体皮肤模型来研究炎症介质对皮肤细胞结构和功能的影响。在本文中,我们描述了两种使用角质形成细胞来研究肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)驱动的皮肤炎症的方案,作为由这些细胞因子驱动的银屑病、白癜风和其他自身免疫性皮肤疾病的替代方法。© 2021 Wiley Periodicals LLC. 基本方案 1:制备 HaCaT 角质形成细胞培养物 基本方案 2:评估 TNF-α 和 IFN-γ 驱动的皮肤炎症的 3D 器官型皮肤培养物。