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A scalable culturing system for the marine annelid Platynereis dumerilii.

PloS one (2019-12-06)
Emily Kuehn, Alexander W Stockinger, Jerome Girard, Florian Raible, B Duygu Özpolat
RÉSUMÉ

Platynereis dumerilii is a marine segmented worm (annelid) with externally fertilized embryos and it can be cultured for the full life cycle in the laboratory. The accessibility of embryos and larvae combined with the breadth of the established molecular and functional techniques has made P. dumerilii an attractive model for studying development, cell lineages, cell type evolution, reproduction, regeneration, the nervous system, and behavior. Traditionally, these worms have been kept in rooms dedicated for their culture. This allows for the regulation of temperature and light cycles, which is critical to synchronizing sexual maturation. However, regulating the conditions of a whole room has limitations, especially if experiments require being able to change culturing conditions. Here we present scalable and flexible culture methods that provide ability to control the environmental conditions, and have a multi-purpose culture space. We provide a closed setup shelving design with proper light conditions necessary for P. dumerilii to mature. We also implemented a standardized method of feeding P. dumerilii cultures with powdered spirulina which relieves the ambiguity associated with using frozen spinach, and helps standardize nutrition conditions across experiments and across different labs. By using these methods, we were able to raise mature P. dumerilii, capable of spawning and producing viable embryos for experimentation and replenishing culture populations. These methods will allow for the further accessibility of P. dumerilii as a model system, and they can be adapted for other aquatic organisms.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Guillard′s (F/2) Marine Water Enrichment Solution, 50 ×, liquid, suitable for plant cell culture
Pyrex® Vacuum filtration system for 47 mm membranes, graduated funnel only
Sigma-Aldrich
Vacuum filtration assembly funnel stems, glass membrane (support), membrane diam. 47 mm, connector for stopper
Millipore
Filtre membrane en esters de cellulose, dimension de pores de 0,22 μm, MF-Millipore, filter diam. 47 mm, hydrophilic, white, Triton-free