title: Culture of neural cells of the eyestalk of a mangrove crab is optimized on poly-L-ornithine substrate.
abstract: Although there is a considerable demand for cell culture protocols from invertebrates for both basic and applied research, few attempts have been made to culture neural cells of crustaceans. We describe an in vitro method that permits the proliferation, growth and characterization of neural cells from the visual system of an adult decapod crustacean. We explain the coating of the culture plates with different adhesive substrates, and the adaptation of the medium to maintain viable neural cells for up to 7 days. Scanning electron microscopy allowed us to monitor the conditioned culture medium to assess cell morphology and cell damage. We quantified cells in the different substrates and performed statistical analyses. Of the most commonly used substrates, poly-L-ornithine was found to be the best for maintaining neural cells for 7 days. We characterized glial cells and neurons, and observed cell proliferation using immunocytochemical reactions with specific markers. This protocol was designed to aid in conducting investigations of adult crustacean neural cells in culture. We believe that an advantage of this method is the potential for adaptation to neural cells from other arthropods and even other groups of invertebrates.
authors.affiliation: Programa de Pós Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.|Programa de Neurobiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho 373, Bloco G2-001, Ilha do Fundão, Rio de Janeiro, RJ, 21949-902, Brazil.
authors.forename: Inês Júlia Ribas
authors.initials: IJ
authors.lastname: Wajsenzon
authors.affiliation: Programa de Pós Graduação em Ciências Biológicas-Fisiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.|Programa de Neurobiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho 373, Bloco G2-001, Ilha do Fundão, Rio de Janeiro, RJ, 21949-902, Brazil.|Neurology Department of Masssachusetts General Hospital, Harvard NeuroDiscovery Center Harvard Medical School, Boston, MA, USA.
authors.forename: Litia Alves
authors.initials: LA
authors.lastname: de Carvalho
authors.affiliation: Universidade Federal do Pará, Campus Universitário do Marajó/Soure, Ilha de Marajó, PA, Brazil.
authors.forename: Adriano
authors.initials: A
authors.lastname: Biancalana
authors.affiliation: Programa de Pós Graduação em Ciências Biológicas-Fisiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.|Programa de Neurobiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho 373, Bloco G2-001, Ilha do Fundão, Rio de Janeiro, RJ, 21949-902, Brazil.
authors.forename: Wagner Antönio Barbosa
authors.initials: WA
authors.lastname: da Silva
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
authors.forename: Claudia
authors.initials: C
authors.lastname: Dos Santos Mermelstein
authors.affiliation: Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil.
authors.forename: Elizabeth Giestal
authors.initials: EG
authors.lastname: de Araujo
authors.affiliation: Programa de Pós Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. sallodi@biof.ufrj.br.|Programa de Pós Graduação em Ciências Biológicas-Fisiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil. sallodi@biof.ufrj.br.|Programa de Neurobiologia, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Av. Carlos Chagas Filho 373, Bloco G2-001, Ilha do Fundão, Rio de Janeiro, RJ, 21949-902, Brazil. sallodi@biof.ufrj.br.
authors.forename: Silvana
authors.initials: S
authors.lastname: Allodi
keywords: Cell culture; Glia; Immunocytochemistry; Invertebrates; Neurons; Visual system