School of Animal and Comparative Biomedical Sciences, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona, United States of America.
BIO5 Institute, University of Arizona, Tucson, Arizona, United States of America.
PLoS One. 2023 Oct 4;18(10):e0292368. doi: 10.1371/journal.pone.0292368. eCollection 2023.
Three-dimensional (3D) culturing techniques can recapitulate the stratified nature of multicellular epithelial tissues. Organotypic 3D epithelial tissue culture methods have several applications, including the study of tissue development and function, drug discovery and toxicity testing, host-pathogen interactions, and the development of tissue-engineered constructs for use in regenerative medicine. We grew 3D organotypic epithelial tissues from foreskin, cervix, and tonsil-derived primary cells and characterized the transcriptome of these in vitro tissue equivalents. Using the same 3D culturing method, all three tissues yielded stratified squamous epithelium, validated histologically using basal and superficial epithelial cell markers. The goal of this study was to use RNA-seq to compare gene expression patterns in these three types of epithelial tissues to gain a better understanding of the molecular mechanisms underlying their function and identify potential therapeutic targets for various diseases. Functional profiling by over-representation and gene set enrichment analysis revealed tissue-specific differences: i.e., cutaneous homeostasis and lipid metabolism in foreskin, extracellular matrix remodeling in cervix, and baseline innate immune differences in tonsil. Specifically, tonsillar epithelia may play an active role in shaping the immune microenvironment of the tonsil balancing inflammation and immune responses in the face of constant exposure to microbial insults. Overall, these data serve as a resource, with gene sets made available for the research community to explore, and as a foundation for understanding the epithelial heterogeneity and how it may impact their in vitro use. An online resource is available to investigate these data (https://viz.datascience.arizona.edu/3DEpiEx/).
三维（3D）培养技术可以再现多细胞上皮组织的分层性质。器官型 3D 上皮组织培养方法有多种应用，包括组织发育和功能研究、药物发现和毒性测试、宿主-病原体相互作用以及用于再生医学的组织工程构建体的开发。我们从包皮、宫颈和扁桃体衍生的原代细胞中培养 3D 器官型上皮组织，并对这些体外组织等效物的转录组进行了表征。使用相同的 3D 培养方法，所有三种组织均产生了分层鳞状上皮，通过基底和表面上皮细胞标志物的组织学验证得到了证实。本研究的目的是使用 RNA-seq 比较这三种上皮组织中的基因表达模式，以更好地了解它们功能背后的分子机制，并确定各种疾病的潜在治疗靶点。过度表达和基因集富集分析的功能分析揭示了组织特异性差异：即包皮中的皮肤稳态和脂质代谢、宫颈中的细胞外基质重塑以及扁桃体中的固有免疫基线差异。具体来说，扁桃体上皮可能在塑造扁桃体的免疫微环境方面发挥积极作用，在不断暴露于微生物侵袭的情况下平衡炎症和免疫反应。总体而言，这些数据可用作资源，提供基因集供研究社区探索，并为理解上皮细胞异质性及其如何影响其体外应用奠定基础。一个在线资源可用于研究这些数据（https://viz.datascience.arizona.edu/3DEpiEx/）。
