Scleractinian coral cell proliferation is reduced in primary culture of suspended multicellular aggregates compared to polyps
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Cell cultures from reef-building scleractinian corals are being developed to study the response of these ecologically important organisms to environmental stress and diseases. Despite the importance of cell division to support propagation, cell proliferation in polyps and in vitro is under-investigated. In this study, suspended multicellular aggregates (tissue balls) were obtained after collagenase dissociation of Pocillopora damicornis coral, with varying yields between enzyme types and brands. Ultrastructure and cell type distribution were characterized in the tissue balls (TBs) compared to the polyp. Morphological evidence of cellular metabolic activity in their ciliated cortex and autophagy in their central mass suggests involvement of active tissue reorganization processes. DNA synthesis was evaluated in the forming multicellular aggregates and in the four cell layers of the polyp, using BrdU labeling of nuclei over a 24 h period. The distribution of BrdU-labeled coral cells was spatially heterogeneous and their proportion was very low in tissue balls (0.2 ± 0.1 %), indicating that suspended multicellular aggregate formation does not involve significant cell division. In polyps, DNA synthesis was significantly lower in the calicoderm (<1 %) compared to both oral and aboral gastroderm (about 10 %) and to the pseudostratified oral epithelium (15–25 % at tip of tentacle). DNA synthesis in the endosymbiotic dinoflagellates dropped in the forming tissue balls (2.7 ± 1.2 %) compared to the polyp (14 ± 3.4 %) where it was not different from the host gastroderm (10.3 ± 1.2 %). A transient (24 h) increase was observed in the cell-specific density of dinoflagellates in individually dissociated coral cell cultures. These results suggest disruption of coral cell proliferation processes upon establishment in primary culture.
KeywordsPrimary culture DNA synthesis BrdU Coral Scleractinia Dinoflagellate Cell proliferation
This work was supported by the ATM Biomineralization of the Museum National d’Histoire Naturelle (‘scleractinian coral’ project) to I. Domart-Coulon, and by European Research Council Advanced Grant 246749 (BIOCARB) to A. Meibom. It was presented at the Symposium on ‘Marine Invertebrate Cell Cultures’, August 30–31 2012, at the Station de Biologie Marine du MNHN in Concarneau, France. We especially thank Michel Hignette and all the team from the Aquarium Tropical, Palais de la Porte Dorée, Paris, France, for access to biological material and facilities for aquarium experiments.
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