Methods for collection, handling, and analysis of sea urchin coelomocytes.
| Autor: | Smith LC; Department of Biological Sciences, George Washington University, Washington, DC, United States. Electronic address: csmith@email.gwu.edu., Hawley TS; National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States., Henson JH; Department of Biology, Dickinson College, Carlisle, PA, United States., Majeske AJ; Department of Biology, University of Puerto Rico, San Juan, Puerto Rico., Oren M; Department of Molecular Biology, Ariel University, Ariel, Israel., Rosental B; Stem Cell Institute, School of Medicine, and the Hopkins Marine Station, Stanford University, Stanford, CA, United States. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in cell biology [Methods Cell Biol] 2019; Vol. 150, pp. 357-389. Date of Electronic Publication: 2019 Jan 09. |
| DOI: | 10.1016/bs.mcb.2018.11.009 |
| Abstrakt: | Sea urchin coelomocytes can be collected in large numbers from adult sea urchins of the species, Strongylocentrotus purpuratus, which typically has 12-40mL of coelomic fluid. Coelomocytes are used for analysis of immune reactions and immune gene expression in addition to basic functions of cells, in particular for understanding structure and modifications of the cytoskeleton in phagocytes. The methods described here include coelomocyte isolation, blocking the clotting reaction, establishing and maintaining primary cultures, separation of different types of coelomocytes into fractions, processing live coelomocytes for light microscopy, fixation and staining for light and electron microscopy, analysis of coelomocyte populations by flow cytometry, and sorting single cells for more detailed follow-up analyses including transcriptomics or genomic characteristics. These methods are provided to make working with coelomocytes accessible to researchers who are unfamiliar with these cells and perhaps to aid others who have worked extensively with invertebrate cells. (© 2019 Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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