Methods for collection, handling, and analysis of sea urchin coelomocytes

L. Courtney Smith, Teresa S. Hawley, John H. Henson, Audrey J. Majeske, Matan Oren, Benyamin Rosental

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

16 Scopus citations

Abstract

Sea urchin coelomocytes can be collected in large numbers from adult sea urchins of the species, Strongylocentrotus purpuratus, which typically has 12–40 mL 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.

Original languageEnglish
Title of host publicationEchinoderms, Part A
PublisherAcademic Press Inc.
Pages357-389
Number of pages33
DOIs
StatePublished - 1 Jan 2019
Externally publishedYes

Publication series

NameMethods in Cell Biology
Volume150
ISSN (Print)0091-679X

Keywords

  • Anticoagulant
  • Coelomocytes
  • Density centrifugation
  • Echinoderm
  • Fixing and staining
  • Flow cytometry
  • Light and electron microscopy
  • Perfusion chamber
  • Primary culture
  • Single cell sorting

ASJC Scopus subject areas

  • Cell Biology

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