Table 2. Summarized advantages and disadvantages of each method tested for coral cell culture.

Method used	Advantages	Disadvantages
Cell dissociation
By simple washing By mechanical scraping with razor blade or scalpel	Easy; Adapted to species with large polyps and/or large tentacles;	Not very effective; Time consuming; yielding ~50%; not adapted to species with small polyps and/or small tentacles;
By paint brush/toothbrush	Relatively easy to perform regardless of the polyp size;	Can be rough for cell membranes and lead to contamination from the mucus layer; time-consuming;
By Ca2+-, Mg2+-free seawater	Simple incubation method;	Effect on desmocytes unknown; mix of single cells and incompletely dissociated tissue fragments detach from the skeleton;
Cell digestion
Enzyme digestion: trypsin	Simple incubation method; high yield; converts proteins to peptides; can promote cell aggregation;	Effect on desmocytes unknown; no standard concentrations or incubation time available; cell clusters; can damage cell surface proteins and their subsequent adhesion capacity; chelating agent necessary to ensure effectiveness (e.g. EDTA);
Enzyme digestion: liberase	Simple incubation method; high yield especially for algae cells;	Effect on desmocytes unknown; no standard concentrations or incubation time available; can affect cell adhesion/aggregation capacity, often presented as a blend of various enzymes;
Cell sorting
Percoll density gradient	Only small doses of Percoll needed each time;	Not cost-effective; density gradients are not always marked enough; protocol must be optimized accordingly to the density characteristics of targeted cells and multiple successive Percoll step gradients needed for full separation;
FACS	Staining process is simple; a lot of data is generated (cell counts per population and relative cell sizes);	Multiple dyes can be costly; partially based on ROS concentration; multiple passes needed for full separation; still new method applied to coral cells;
Growth medium	Methods established with other living organisms; many products available to purchase;	Does not correspond exactly to coral cell needs; must be combined with additives for optimization;
DMEM
RPMI
F12
FBS
Antibiotic treatment
Pen-Strep (Penicillin and Streptomycin)	Method established with other living organisms; many products available to purchase;	Little effect on the bacterial and viral population observed;
Anti-Anti (Penicillin, Streptomycin, Amphotericin B);	Method established with other living organisms; many products available to purchase; More effective than Pen-Strep alone;
Gentamicin	Method established with other living organisms; many products available to purchase;	Most efficient when combined with other antibiotics that target DNA synthesis;
Cell attachment
Untreated glass or plastic	Simple;	~40% of cells lost;
Collagen coating (glass or plastic)	Can be tested with macromolecules other than collagen;	Might need to be tailored to different cell types;
Cell visualization for counting
Trypan Blue	No incubation necessary;	Lethal stain; reacts with salts and creates clusters;