ABSTRACT
The sea urchin is an emergent model system for studying basic and translational immunology. Here we report a new method for the harvesting and maintenance of primary immune cells isolated from adult Paracentrotus lividus, a common Mediterranean sea urchin species. This optimised method uses coelomocyte culture medium, containing a high-affinity Ca2+ chelator, as the ideal harvesting and anti-clotting vehicle and short-term culture medium (≤48 h), and artificial seawater as the master medium that maintains cell survival and in vitro-ex vivo physiological homeostasis over 2 weeks. Gradually reducing the amount of anticoagulant solution in the medium and regularly replacing the medium led to improved culture viability. Access to a robust and straightforward in vitro-ex vivo system will expedite our understanding of deuterostome immunity as well as underscore the potential of sea urchin with respect to biomedicine and regulatory testing.
This article has an associated First Person interview with the first author of the paper.
Footnotes
Competing interests
The authors declare no competing or financial interests.
Author contributions
Conceptualization: A.P.; Methodology: A.P., A.A.; Validation: A.P., A.A.; Formal analysis: A.A.; Investigation: A.P., A.A.; Data curation: A.P., A.A.; Writing - original draft: A.P., A.A.; Writing - review & editing: A.P.; Supervision: A.P.; Project administration: A.P.; Funding acquisition: A.P.
Funding
This project has received funding from the European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement [No 671881].
Supplementary information
Supplementary information available online at http://bio.biologists.org/lookup/doi/10.1242/bio.039289.supplemental
- Received October 12, 2018.
- Accepted January 29, 2019.
- © 2019. Published by The Company of Biologists Ltd
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
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