POL Scientific / JBM / Volume 5 / Issue 4 / DOI: 10.14440/jbm.2018.237
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PROTOCOLS

Method for selective quantification of immune and inflammatory cells in the cornea using flow cytometry

Mamoru Ogawa1,2 Takenori Inomata3,4 Tina Shiang5 Kazuo Tsubota2 Akira Murakami3
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1 Laboratory for Metabolomics, RIKEN Center for Integrative Medical Sciences, Tsurumi-ku, Yokohama 230-0045, Japan
2 Department of Ophthalmology, Keio University School of Medicine, Tokyo 160-8582, Japan
3 Department of Ophthalmology, Juntendo University Faculty of Medicine, Tokyo, 113-0033, Japan
4 Department of Strategic Operating Room Management and Improvement, Juntendo University Faculty of Medicine, Tokyo, 113-0033, Japan
5 University of Massachusetts Medical School, Department of Radiology, MA 01655, USA
Published: 22 November 2018
© 2018 by the author. Licensee POL Scientific, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

The cornea serves as a protective surface against the environment (i.e., 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 in vivo.

Keywords
cornea
flow cytometry
angiogenesis
inflammation
immune privilege
transplantation
wound healing
References

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Journal of Biological Methods, Electronic ISSN: 2326-9901 Print ISSN: TBA, Published by POL Scientific