Method for selective quantification of immune and inflammatory cells in the cornea using flow cytometry
Method for selective quantification of immune and inflammatory cells in the cornea using flow cytometry
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DOI：
10.14440/jbm.2018.237
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Murakami, Akira
中科院分区：
文献类型：
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作者：
Ogawa, Mamoru;Inomata, Takenori;Murakami, Akira
角膜是抵御环境（即过敏原、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。该协议描述了流式细胞术的角膜准备，以评估角膜中的局部细胞。我们的模型详细介绍了整个过程，从确定实验中需要多少角膜到角膜切除，再到角膜细胞的消化和染色。该模型的简单性允许系统分析不同的角膜免疫、炎症、血管生成和伤口愈合机制。在角膜移植中，驻留免疫和炎症细胞是血管生成、混浊和移植排斥机制的关键。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，该模型可用于分析滴注和结膜下注射等新药物的功效，并评估治疗分子在体内增强移植物存活和伤口愈合的潜力。
The cornea serves as a protective surface against the environment (i.e., allergens, pollutants, desiccation and micro-organisms) 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 in vivo.