Shannon Jessica L, Kirchner Stephen J, Zhang Jennifer Y
Department of Dermatology, Duke University, Durham NC, USA.
Bio Protoc. 2022 Sep 20;12(18). doi: 10.21769/BioProtoc.4514.
The experimentation with surgically discarded human skin represents a unique methodology amenable for mechanism and pharmacologic agent studies without the involvement of human subjects. Here, we describe a protocol that includes preparation, culture, and stimulation of human skin explants, and subsequent analyses by quantitative reverse transcription PCR and immunostaining. This protocol may also be applied for studies of murine skin, reducing animal numbers and potentially harmful treatments. In our hands, this protocol has been used for wound healing, viral infection, and hair growth-related studies. Graphical abstract: Cartoon of explant skin culture. Skin explant sits on top of a gelatin surgical sponge saturated with culture medium at an air-liquid interface.
利用手术废弃的人体皮肤进行实验是一种独特的方法，适用于机制和药物研究，且无需涉及人体受试者。在此，我们描述了一种方案，包括人体皮肤外植体的制备、培养和刺激，以及随后通过定量逆转录聚合酶链反应和免疫染色进行的分析。该方案也可应用于小鼠皮肤研究，减少动物数量和潜在的有害处理。在我们的实践中，该方案已用于伤口愈合、病毒感染和毛发生长相关研究。图形摘要：外植体皮肤培养示意图。皮肤外植体置于气液界面处充满培养基的明胶手术海绵上。
