Department of Otorhinolaryngology: Head and Neck Surgery, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
Department of Dermatology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
STAR Protoc. 2023 Mar 17;4(1):102101. doi: 10.1016/j.xpro.2023.102101. Epub 2023 Feb 6.
Here, we provide a protocol to model the effects of changes to a small number of cells, such as those arising from a mutation or a virus infection, in stratified epithelia. We describe steps for diluting engineered human keratinocytes into a larger population of unmodified cells and using these cells to grow three-dimensional organotypic cultures. We detail steps to observe effects that are not apparent in homogenous organotypic epithelial cultures by visualizing the localization of modified keratinocytes in epithelial layers. For complete details on the use and execution of this protocol, please refer to Hatterschide et al. (2022)..
在这里，我们提供了一个方案来模拟少数细胞变化的影响，例如由突变或病毒感染引起的变化，在分层上皮组织中。我们描述了将工程化的人类角质形成细胞稀释到更大数量的未修饰细胞中，并使用这些细胞来生长三维器官型培养物的步骤。我们详细介绍了通过观察修饰的角质形成细胞在上皮层中的定位来观察均质上皮培养物中不明显的效果的步骤。有关该方案的使用和执行的完整详细信息，请参阅 Hatterschide 等人。(2022)。
