Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies

Detalhes bibliográficos
Autor(a) principal: Lampreia, Fábio Gabriel Pereira
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/10868
Resumo: Dissertação para obtenção do Grau de Mestre em Biotecnologia
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spelling Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategiesNeural stem cellsBrain derived neurotrophic factorNon-viral gene deliveryLipofectionPlasmid DNANeurotoxicityDissertação para obtenção do Grau de Mestre em BiotecnologiaThe brain derived neurotrophic factor holds neuroprotective and neurogenic roles. The presence of this factor is believed to present highly beneficial effects on injured cells within several neurological disorders, where the levels of this neurotrophin are usually drastically decreased. Neural stem cells are multipotent, self-renewing cells with the ability to differentiate into the three main type of cells within the central nervous system - neurons, astrocytes and oligodendrocytes. The combination of gene therapy, through the introduction of therapeutic genes into the desired cells, with cell therapy aiming for the replacement of damaged cells within a given disorder, hold great promise in modern regenerative medicine. The aim of this work was the overexpression of the brain derived neurotrophic factor in neural stem cells. For this, mouse and human neural stem cells were transfected with one of three non-viral techniques - microporation, lipid and cationic-polymer based strategies. Human NSC were efficiently transfected using the commercial lipid-based transfection reagent Lipofectamine 2000 with an efficiency of 35%, maintaining their differentiation potential. Cells within the differentiation process were efficiently transfected with 13% efficiency. Cell viability has always remained above 70% after the lipofection process. The transfection with the BDNF gene resulted in neurons with longer primary neurites, and more secondary neurites than control cells, which hints at the promotion of neurite outgrowth and ramification of neurons by this neurotrophin. Finally, healthy cells were exposed to toxic concentrations of glutamate. Conditioned media containing secreted BDNF from transfected cells was able to protect these cells from glutamate-induced neurotoxicity, as well as reducing the levels of expression of the pro-apoptotic protein caspase7 to near-control levels. Overall, this work provides the first evidences of the successful use of BDNF-overexpressing NSC, based on a non-viral gene delivery approach for decreasing neurotoxicity.Faculdade de Ciências e TecnologiaMadeira, TeresaRUNLampreia, Fábio Gabriel Pereira2013-12-16T11:53:50Z20132013-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/10868TID:201755238enginfo: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:RCAAP2024-03-11T03:44:53Zoai:run.unl.pt:10362/10868Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:19:47.891306Repositório 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 Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
title Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
spellingShingle Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
Lampreia, Fábio Gabriel Pereira
Neural stem cells
Brain derived neurotrophic factor
Non-viral gene delivery
Lipofection
Plasmid DNA
Neurotoxicity
title_short Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
title_full Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
title_fullStr Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
title_full_unstemmed Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
title_sort Overexpressing BDNF in Neural Stem Cells using non-viral gene delivery strategies
author Lampreia, Fábio Gabriel Pereira
author_facet Lampreia, Fábio Gabriel Pereira
author_role author
dc.contributor.none.fl_str_mv Madeira, Teresa
RUN
dc.contributor.author.fl_str_mv Lampreia, Fábio Gabriel Pereira
dc.subject.por.fl_str_mv Neural stem cells
Brain derived neurotrophic factor
Non-viral gene delivery
Lipofection
Plasmid DNA
Neurotoxicity
topic Neural stem cells
Brain derived neurotrophic factor
Non-viral gene delivery
Lipofection
Plasmid DNA
Neurotoxicity
description Dissertação para obtenção do Grau de Mestre em Biotecnologia
publishDate 2013
dc.date.none.fl_str_mv 2013-12-16T11:53:50Z
2013
2013-01-01T00:00:00Z
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/10362/10868
TID:201755238
url http://hdl.handle.net/10362/10868
identifier_str_mv TID:201755238
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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