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Long-Term Culture of Mouse EmbryonicStem Cell-Derived Adherent Neurospheres and Functional Neurons
Hayashi, Mirian A. F; Guerreiro, Juliano R; Cassola, Antonio C; Lizier, Nelson F; Kerkis, Alexandre; Camargo, Antonio C. M; Kerkis, Irina.
Afiliação
  • Hayashi, Mirian A. F; Instituto Butantan. São Paulo. BR
  • Guerreiro, Juliano R; Instituto Butantan. São Paulo. BR
  • Cassola, Antonio C; s.af
  • Lizier, Nelson F; Instituto Butantan. São Paulo. BR
  • Kerkis, Alexandre; s.af
  • Camargo, Antonio C. M; Instituto Butantan. São Paulo. BR
  • Kerkis, Irina; Instituto Butantan. São Paulo. BR
Tissue Engineering Part C Methods ; 16(6): 1493-1502, Dec.2010.
Article em En | SES-SP, SESSP-IBPROD, SES-SP, SESSP-IBACERVO | ID: biblio-1068173
Biblioteca responsável: BR78.1
Localização: BR78.1
ABSTRACT
Innumerous protocols, using the mouse embryonic stem (ES) cells as model for in vitro study of neuronsfunctional properties and features, have been developed. Most of these protocols are short lasting, which,therefore, does not allow a careful analysis of the neurons maturation, aging, and death processes. We describehere a novel and efficient long-lasting protocol for in vitro ES cells differentiation into neuronal cells. It consists of obtaining embryoid bodies, followed by induction of neuronal differentiation with retinoic acid of nonadherent embryoid bodies (three-dimensional model), which further allows their adherence and formation of adherent neurospheres (AN, bi-dimensional model). The AN can be maintained for at least 12 weeks in culture under repetitive mechanical splitting, providing a constant microenvironment (in vitro niche) for the neuronal progenitor cells avoiding mechanical dissociation of AN. The expression of neuron-specific proteins, such as nestin, sox1, beta III-tubulin, microtubule-associated protein 2, neurofilament medium protein, Tau, neuronal nuclei marker, gamma-aminobutyric acid, and 5-hydroxytryptamine, were confirmed in these cells maintained during3 months under several splitting...
Assuntos
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Coleções: 06-national / BR Base de dados: SES-SP / SESSP-IBACERVO / SESSP-IBPROD Assunto principal: Biomarcadores / Diferenciação Celular Limite: Animals Idioma: En Revista: Tissue Engineering Part C Methods Ano de publicação: 2010 Tipo de documento: Article
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Coleções: 06-national / BR Base de dados: SES-SP / SESSP-IBACERVO / SESSP-IBPROD Assunto principal: Biomarcadores / Diferenciação Celular Limite: Animals Idioma: En Revista: Tissue Engineering Part C Methods Ano de publicação: 2010 Tipo de documento: Article