Development of novel human cellular models for neurotoxicity studies

Detalhes bibliográficos
Autor(a) principal: Terrasso, Ana Paula Barreto
Data de Publicação: 2012
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/8488
Resumo: Dissertation to obtain master degree in Genética Molecular e Biomedicina
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spelling Development of novel human cellular models for neurotoxicity studiesHuman stem cellsNTera2/cloneD1 cell lineNeural differentiationStirred suspension culture systems3D cell modelsAggregatesDissertation to obtain master degree in Genética Molecular e BiomedicinaInformation currently available on neurotoxicity of chemicals is scarce and there are a growing number of new compounds to be tested. Therefore, new strategies are necessary to identify neurotoxic agents with speed, reliability and respect for animal welfare. The limited availability of primary human brain cells means that there is a need for human cell lines that reliably model human neurons and astrocytes. Despite the advances in stem cell research, numerous challenges must be overcome before this technology can be widespread used, such as low differentiation efficiency. Human pluripotent embryocarcinoma NTera2/cloneD1 (NT2) cell line is an alternative cell source from which neurons and astrocytes can be derived in vitro. The aim of this work was to develop scalable and reproducible novel human cellular models using NT2 cells as source of differentiated neural phenotypes. A 2D culture system for astrocytic differentiation was implemented. After 4 weeks of differentiation with retinoic acid followed by 5 weeks maturation with mitotic inhibitors, astrocytes obtained expressed vimentin, GFAP, S100- and GLT-1 as characterized by immunodetection and qRT-PCR. Then, a 3D culture approach was adopted, using stirred suspension culture systems, in which cell-cell and cell-extracellular matrix interactions occur, mimicking better the in vivo situation. NT2 cells, inoculated as single cells, spontaneously aggregated without compromising their pluripotency. Optimization of stirring rate allowed control of aggregate size along time. After 3 weeks of RA treatment and 2 weeks of maturation, neurons expressing βIII-tubulin, MAPs and synaptophysin and astrocytes expressing vimentin, GFAP, S100- and GLT-1 were detected, as characterized by immunodetection and qRT-PCR. Furthermore, astrocytes presented a 2.5-fold higher yield than that observed in 2D culture systems. Results showed that NT2 differentiated cells are promising models for neurotoxicity testing. Furthermore, the 3D culture systems developed herein can contribute to increase the relevance of these studies, recapitulating human neuron-astrocyte interactions in a 3D cellular context.Fundação para a Ciência e Tecnologia - PTDC/EEB-BIO/112786/2009Faculdade de Ciências e TecnologiaBrito, CatarinaCaldas, MargaridaRUNTerrasso, Ana Paula Barreto2013-01-10T10:16:26Z20122012-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/8488enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T03:41:08Zoai:run.unl.pt:10362/8488Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:18:13.791774Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Development of novel human cellular models for neurotoxicity studies
title Development of novel human cellular models for neurotoxicity studies
spellingShingle Development of novel human cellular models for neurotoxicity studies
Terrasso, Ana Paula Barreto
Human stem cells
NTera2/cloneD1 cell line
Neural differentiation
Stirred suspension culture systems
3D cell models
Aggregates
title_short Development of novel human cellular models for neurotoxicity studies
title_full Development of novel human cellular models for neurotoxicity studies
title_fullStr Development of novel human cellular models for neurotoxicity studies
title_full_unstemmed Development of novel human cellular models for neurotoxicity studies
title_sort Development of novel human cellular models for neurotoxicity studies
author Terrasso, Ana Paula Barreto
author_facet Terrasso, Ana Paula Barreto
author_role author
dc.contributor.none.fl_str_mv Brito, Catarina
Caldas, Margarida
RUN
dc.contributor.author.fl_str_mv Terrasso, Ana Paula Barreto
dc.subject.por.fl_str_mv Human stem cells
NTera2/cloneD1 cell line
Neural differentiation
Stirred suspension culture systems
3D cell models
Aggregates
topic Human stem cells
NTera2/cloneD1 cell line
Neural differentiation
Stirred suspension culture systems
3D cell models
Aggregates
description Dissertation to obtain master degree in Genética Molecular e Biomedicina
publishDate 2012
dc.date.none.fl_str_mv 2012
2012-01-01T00:00:00Z
2013-01-10T10:16:26Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/8488
url http://hdl.handle.net/10362/8488
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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