Ogawa Mamoru, Inomata Takenori, Shiang Tina, Tsubota Kazuo, Murakami Akira
Laboratory for Metabolomics, RIKEN Center for Integrative Medical Sciences, Tsurumi-ku, Yokohama 230-0045, Japan.
Department of Ophthalmology, Keio University School of Medicine, Tokyo 160-8582, Japan.
J Biol Methods. 2018 Nov 22;5(4):e102. doi: 10.14440/jbm.2018.237. eCollection 2018.
The cornea serves as a protective surface against the environment (, allergens, pollutants, desiccation and microorganisms) and promotes vision, made possible by corneal transparency. This protocol describes corneal preparation for flow cytometry to assess cells localized in the cornea. Our model details the process, from determining how many corneas are needed in the experiment to corneal excision to digestion and staining of the cornea cells. The simplicity of the model allows for systematic analysis of different corneal mechanisms of immunity, inflammation, angiogenesis and wound healing. In corneal transplantation, residential immune and inflammatory cells are key to the mechanisms that underlie angiogenesis, opacity, and graft rejection. In addition, this model can also elucidate cellular mechanisms mediating corneal graft outcomes and wound healing. Lastly, this model can be used to analyze the efficacy of new medications such as instillation and subconjunctival injections and assess the potential of therapeutic molecules to enhance graft survival and wound healing .
角膜作为抵御外界环境（过敏原、污染物、干燥和微生物）的保护表面，并通过角膜透明度促进视力。本方案描述了用于流式细胞术的角膜制备方法，以评估角膜中的局部细胞。我们的模型详细介绍了该过程，从确定实验所需角膜的数量到角膜切除，再到角膜细胞的消化和染色。该模型的简单性使得能够系统地分析角膜免疫、炎症、血管生成和伤口愈合的不同机制。在角膜移植中，驻留免疫细胞和炎症细胞是血管生成、角膜混浊和移植排斥反应潜在机制的关键。此外，该模型还可以阐明介导角膜移植结果和伤口愈合的细胞机制。最后，该模型可用于分析新药物（如滴眼剂和结膜下注射）的疗效，并评估治疗分子提高移植存活率和促进伤口愈合的潜力。
