Sea urchin Paracentrotus lividus immune cells in culture: formulation of the appropriate harvesting and culture media and maintenance conditions.

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.