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Cell tracking and velocimetric parameters analysis as an approach to assess activity of mussel (Mytilus edulis) hemocytes in vitro

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Abstract

Hemocytes constitute the key element of innate immunity in bivalves, being responsible for secretion of antimicrobial peptides and release of zymogens from the prophenoloxidase system within the hemolymph compartment, reactive oxygen species production and phagocytosis. Hemocytes are found (and collected) as cells in suspension in circulating hemolymph. Hemocytes are adherent cells as well, infiltrating tissues and migrating to infected areas. In the present study, we applied an approach based on fluorescent staining and nuclei-tracking to determine migration velocity of hemocytes from the blue mussel, Mytilus edulis, in culture. Freshly collected hemocytes attached to substrate and start to move spontaneously in few minutes. Two main hemocyte morphologies can be observed: small star-shaped cells which were less motile and spread granular cells with faster migrations. Cell-tracking was combined to MTT mitochondria metabolic rate measurements in order to monitor global cell population activity over 4 days of culture. A transient peak of cell activity was recorded after 24–48 h of culture, corresponding to a speed up of cell migration. Videomicroscopy and cell tracking techniques provide new tools to characterize activity of mussel immunocytes in culture. Our analysis of hemocyte migration reveals that motility is very sensitive to cell environmental factors.

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Acknowledgments

This work was supported by grants from the State/Region Plan Contract (CPER) allocated through the Research Federation FED 4116 SCALE (Sciences Appliquées à L’Environnement). Damien Rioult were recipients for doctoral fellowships from the Conseil Regional de Haute-Normandie.

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Authors and Affiliations

  1. Laboratory of Ecotoxicology, EA 3222, FED 4116 SCALE, University of Le Havre, 76058, Le Havre Cedex, France

    Damien Rioult & Frank Le Foll

  2. CNRS INEE, FRE3484 BioMEA Biologie des Mollusques marins et des Ecosystèmes Associés, IBFA, University of Caen, 14032, Caen Cedex, France

    Jean-Marc Lebel

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  1. Damien Rioult
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  2. Jean-Marc Lebel
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  3. Frank Le Foll
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Correspondence to Damien Rioult.

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Rioult, D., Lebel, JM. & Le Foll, F. Cell tracking and velocimetric parameters analysis as an approach to assess activity of mussel (Mytilus edulis) hemocytes in vitro. Cytotechnology 65, 749–758 (2013). https://doi.org/10.1007/s10616-013-9558-2

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