Johansen Claus
Department of Dermatology, Aarhus University Hospital;
J Vis Exp. 2017 Dec 22(130):56863. doi: 10.3791/56863.
The main function of keratinocytes is to provide the structural integrity of the epidermis, thereby maintaining a mechanical barrier to the outside world. In addition, keratinocytes play an essential role in the initiation, maintenance, and regulation of epidermal immune responses by being part of the innate immune system responding to antigenic stimuli in a fast, nonspecific manner. Here, we describe a protocol for isolation of primary human keratinocytes from adult skin, and demonstrate that these cells respond to calcium-induced terminal differentiation, as measured by an increased expression of the differentiation marker involucrin. In addition, we show that the isolated keratinocytes are responsive to IL-1β-induced activation of intracellular signaling pathways as measured by the activation of the p38 MAPK pathway. Taken together, we describe a method for isolation and culturing of primary human keratinocytes from adult skin. Because the keratinocytes are the predominant cell type in the epidermis, this method is useful to study molecular mechanisms in cutaneous biology in vitro.
角质形成细胞的主要功能是提供表皮的结构完整性，从而维持对外界的机械屏障。此外，角质形成细胞作为先天免疫系统的一部分，以快速、非特异性的方式对抗抗原刺激，在表皮免疫反应的启动、维持和调节中发挥着重要作用。在此，我们描述了一种从成人皮肤中分离原代人角质形成细胞的方案，并证明这些细胞对钙诱导的终末分化有反应，这通过分化标志物兜甲蛋白表达的增加来衡量。此外，我们表明，通过p38丝裂原活化蛋白激酶（MAPK）途径的激活来衡量，分离的角质形成细胞对白细胞介素-1β（IL-1β）诱导的细胞内信号通路激活有反应。综上所述，我们描述了一种从成人皮肤中分离和培养原代人角质形成细胞的方法。由于角质形成细胞是表皮中的主要细胞类型，该方法有助于在体外研究皮肤生物学中的分子机制。
