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Culture of neural cells of the eyestalk of a mangrove crab is optimized on poly-l-ornithine substrate

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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.

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Acknowledgments

We thank Sergio Luiz de Carvalho for the advice on the figures. We are grateful to the Multi-user Unit of Image of the Instituto de Biofísica Carlos Chagas Filho of the Universidade Federal do Rio de Janeiro for the use of the Zeiss Axio Imager M2 inverted fluorescent microscope. The authors are also indebted to the Rudolf Barth Electron Microscopy Platform of the Oswaldo Cruz Institute/Fiocruz. This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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  1. Litia Alves de Carvalho

    Present address: Neurology Department of Masssachusetts General Hospital, Harvard NeuroDiscovery Center Harvard Medical School, Boston, MA, USA

Authors and Affiliations

  1. Programa de Pós Graduação em Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Inês Júlia Ribas Wajsenzon & Silvana Allodi

  2. 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

    Litia Alves de Carvalho, Wagner Antönio Barbosa da Silva & Silvana Allodi

  3. Universidade Federal do Pará, Campus Universitário do Marajó/Soure, Ilha de Marajó, PA, Brazil

    Adriano Biancalana

  4. Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Claudia dos Santos Mermelstein

  5. Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil

    Elizabeth Giestal de Araujo

  6. 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

    Inês Júlia Ribas Wajsenzon, Litia Alves de Carvalho, Wagner Antönio Barbosa da Silva & Silvana Allodi

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  1. Inês Júlia Ribas Wajsenzon
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Correspondence to Silvana Allodi.

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Wajsenzon, I.J.R., de Carvalho, L.A., Biancalana, A. et al. Culture of neural cells of the eyestalk of a mangrove crab is optimized on poly-l-ornithine substrate. Cytotechnology 68, 2193–2206 (2016). https://doi.org/10.1007/s10616-015-9942-1

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