title: Development of long-term primary cell aggregates from Mediterranean octocorals.
abstract: In lower metazoans, the aggregative properties of dissociated cells leading to in vitro stable multicellular aggregates have furnished a remarkable experimental material to carry out investigations in various research fields. One of the main expectations is to find good models for the study in vitro of the first steps of biomineralization processes. In this study, we examined five common Mediterranean gorgonians (Paramuricea clavata, Corallium rubrum, Eunicella singularis, Eunicella cavolinii, and Eunicella verrucosa) using mechanical cell aggregate production techniques. In particular, we investigated the conditions of aggregate formation, their number and survival in experimental conditions, the DNA synthesizing activity using 5'-bromo-2'-deoxyuridine (BrdU) tests, and the response to calcein addition and observed the secretion of newly formed sclerites. The BrdU tests showed that cell proliferation depends on the size of aggregates and on the presence/absence of symbiotic zooxanthellae. With epifluorescent and confocal imaging from calcein addition assays, we observed the presence of calcium ions within cells, a possible clue for prediction of sclerite formation or calcium deposition. The species-specific efficiency in production of cell aggregates is correlated to the size of polyps, showing that the higher density of polyps and their diameter correspond to higher production of cell aggregates. Regarding the long-term maintenance, we obtained the best results from E. singularis, which formed multicellular aggregates of 0.245 mm ± 0.086 mm in size and maintained symbiotic association with zooxanthellae throughout the experimental run. Formation of sclerites within aggregates opens a wide field of investigation on biomineralization, since de novo sclerites were observed around 30 d after the beginning of the experiment.
authors: Dipartimento di Scienze della Vita e dell'Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131, Ancona, Italy.
authors: Carla
authors: C
authors: Huete-Stauffer
authors: Dipartimento di Scienze della Vita e dell'Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131, Ancona, Italy.
authors: Laura
authors: L
authors: Valisano
authors: Dipartimento di Scienze della Terra, dell'Ambiente e della Vita (DISTAV), Università degli Studi di Genova, Corso Europa 26, 16132, Genoa, Italy.
authors: Elda
authors: E
authors: Gaino
authors: Dipartimento di Scienze della Terra, dell'Ambiente e della Vita (DISTAV), Università degli Studi di Genova, Corso Europa 26, 16132, Genoa, Italy.
authors: Luigi
authors: L
authors: Vezzulli
authors: Dipartimento di Scienze della Vita e dell'Ambiente (DiSVA), Università Politecnica delle Marche, Via Brecce Bianche, 60131, Ancona, Italy. c.cerrano@univpm.it.
authors: Carlo
authors: C
authors: Cerrano
mesh_terms: D000818:Animals; D037421:Anthozoa / Q000166:cytology* / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D002449:Cell Aggregation / Q000502:physiology*; D002470:Cell Survival; D004247:DNA / Q000096:biosynthesis; D046529:Microscopy, Electron, Transmission
chemical_list: D004247:DNA
keywords: Biomineralization; Calcium; Cell culture; Octocorals; Zooxanthellae