title: Isolation and regeneration of protoplasts from Ectocarpus.
abstract: This article describes how to obtain isolated cells with no surrounding cell wall by enzymatic digestion of Ectocarpus filaments. The resultant protoplasts are totipotent and regenerate to produce individual algae under appropriate culture conditions. The yield of protoplasts and their capacity to regenerate are highly dependent on the Ectocarpus strain used, the stage of the life cycle, and the culture conditions. The highest yields are obtained with young gametophyte filaments cultivated at low density. The naked, wall-less cells produced by this protocol can be used for several applications, including studies of cell wall regeneration, investigation of the role of the cell wall in determining cell fate, and as a source of naked cells for the development of methods for introducing diverse molecules into the cell.
authors.affiliation: UPMC Université Paris 06, The Marine Plants and Biomolecules Laboratory, UMR 7139, Station Biologique de Roscoff, 29682 Roscoff Cedex, France.
authors.forename: Susana M
authors.initials: SM
authors.lastname: Coelho
authors.forename: Delphine
authors.initials: D
authors.lastname: Scornet
authors.forename: Sylvie
authors.initials: S
authors.lastname: Rousvoal
authors.forename: Nick
authors.initials: N
authors.lastname: Peters
authors.forename: Laurence
authors.initials: L
authors.lastname: Dartevelle
authors.forename: Akira F
authors.initials: AF
authors.lastname: Peters
authors.forename: J Mark
authors.initials: JM
authors.lastname: Cock
mesh_terms: D003584:Cytological Techniques / Q000379:methods*; D000459:Phaeophyceae / Q000166:cytology*; D011523:Protoplasts / Q000502:physiology*