Rana Isha, Badarinath Krithika, Zirmire Ravindra K, Jamora Colin
IFOM-inStem Joint Research Laboratory, Centre for Inflammation and Tissue Homeostasis, Institute for Stem Cell Science and Regenerative Medicine (inStem), Bangalore, Karnataka, India.
Shanmugha Arts, Science, Technology and Research Academy (SASTRA) University, Thanjavur, Tamil Nadu, India.
Bio Protoc. 2021 Sep 5;11(17):e4148. doi: 10.21769/BioProtoc.4148.
The skin plays an important role in protecting the body from pathogens and chemicals in the external environment. Upon injury, a healing program is rapidly initiated and involves extensive intercellular communication to restore tissue homeostasis. The deregulation of this crosstalk can lead to abnormal healing processes and is the foundation of many skin diseases. A relatively overlooked cell type that nevertheless plays critical roles in skin homeostasis, wound repair, and disease is the dendritic epidermal T cells (DETCs), which are also called γδT-cells. Given their varied roles in both physiological and pathological scenarios, interest in the regulation and function of DETCs has substantially increased. Moreover, their ability to regulate other immune cells has garnered substantial attention for their potential role as immunomodulators and in immunotherapies. In this article, we describe a protocol to isolate and culture DETCs and analyse them within the skin. These approaches will facilitate the investigation of their crosstalk with other cutaneous cells and the mechanisms by which they influence the status of the skin. Graphic abstract: Overall workflow to analyse DETCs and .
皮肤在保护身体免受外部环境中的病原体和化学物质侵害方面发挥着重要作用。受伤后,愈合程序会迅速启动,涉及广泛的细胞间通讯以恢复组织稳态。这种相互作用的失调会导致异常的愈合过程,是许多皮肤病的基础。一种相对被忽视但在皮肤稳态、伤口修复和疾病中起关键作用的细胞类型是树突状表皮T细胞(DETC),也称为γδT细胞。鉴于它们在生理和病理情况下的多种作用,对DETC调节和功能的兴趣大幅增加。此外,它们调节其他免疫细胞的能力因其作为免疫调节剂和在免疫疗法中的潜在作用而备受关注。在本文中,我们描述了一种分离和培养DETC并在皮肤内对其进行分析的方案。这些方法将有助于研究它们与其他皮肤细胞的相互作用以及它们影响皮肤状态的机制。图形摘要:分析DETC的总体工作流程。