Division of Dermatology, Department of Internal Related, Kobe University Graduate School of Medicine, Kobe, Japan.
Department of Dermatology, Keio University School of Medicine, Tokyo, Japan.
Curr Protoc. 2022 Jul;2(7):e464. doi: 10.1002/cpz1.464.
The epidermis is a stratified epithelium. Compared to that for monolayered epithelia, understanding of the cell biology of stratified epithelia lags far behind. The major reason for this is the limitation of methods to reproduce the epidermis in vitro using cultured keratinocytes: for example, cultured keratinocyte cell sheets lack Langerhans cells, melanocytes, nerves, sweat ducts, and hair follicles. One current way to overcome this limitation is to observe the epidermis in vivo via whole-mount staining and three-dimensional imaging. Here, we describe how to prepare epidermal sheets from skin and how to immunostain and observe them in whole mount. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Preparation of mouse epidermal sheets by the ammonium thiocyanate method Alternate Protocol: Preparation of mouse epidermal sheets by the dispase method Basic Protocol 2: Preparation of human epidermal sheets by the dispase method Basic Protocol 3: Whole-mount immunostaining of epidermis.
表皮是一种复层上皮。与单层上皮相比，人们对复层上皮的细胞生物学的了解还远远落后。造成这种情况的主要原因是使用培养的角质形成细胞在体外再现表皮的方法有限：例如，培养的角质形成细胞片层缺乏朗格汉斯细胞、黑色素细胞、神经、汗腺和毛囊。目前克服这一限制的一种方法是通过全染色和三维成像来观察体内的表皮。在这里，我们描述了如何从皮肤中制备表皮片以及如何在全染色中进行免疫染色和观察。© 2022 威利父子公司。基本方案 1：用硫氰酸铵法制备小鼠表皮片替代方案：用Dispase 法制备小鼠表皮片基本方案 2：用 Dispase 法制备人表皮片基本方案 3：表皮的全染色免疫组织化学
