Establishing a cell biology platform: isolation and preservation of human blood products

Detalhes bibliográficos
Autor(a) principal: Marques, Graça Susete Costa de Carvalho
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/11009
Resumo: Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina
id RCAP_ed3f5cfbd032612c7584df57ae269dcb
oai_identifier_str oai:run.unl.pt:10362/11009
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling Establishing a cell biology platform: isolation and preservation of human blood productsSeraCell isolationCryopreservationCell culturePBMCs (peripheral blood mononuclear cells)Dissertação para obtenção do Grau de Mestre em Genética Molecular e BiomedicinaThe use of human primary cells provide researchers in different areas with irrefutable more biologically relevant data than using cell lines or animal blood cells. The work was performed in the scope of the Cell Biology Services @ CEDOC, aiming to provide viable and trustful human primary cells and products. We had three main objectives: protocol optimizations for blood cell isolation, culture and cryopreservation; cost estimation and divulgation of the services. We have reviewed standard protocols and compared different strategies for blood cell isolation. The impact of those methodologies was evaluated regarding cell yield and purity, cell functional characteristics and cost. We also developed a method for serum isolation from human plasma in blood buffy coats. The resultant sera were sterile and suitable to be used in leukocyte cultures. Different protocols for T cells isolation were compared: positive versus negative immunomagnetic selection and isolation using nylon wool fiber columns. Positive selection provided the highest isolation yield (32.35%), while negatively selected cells had the highest purity (92.81%). Although nylon wool fiber column was the fastest and cheapest method, unlike the immunomagnetic methods, it did not allow complete separation of T from B lymphocytes. Positive selection of monocytes was compared using two widely used commercial kits. Miltenyi’s kit provided the highest isolation yield (25.92%), recovery rate (86.70%) and purity (95.01%). Monocytes isolated with StemCell kit presented a higher cell complexity, and when differentiated into dendritic cells (DCs), showed a more mature phenotype. Differences between both kits are probably caused by the nature of the magnetic beads, suggesting caution when choosing one or other kit, as it may have an impact on DCs’ function. Overall, although dealing with apparently straight forward methodologies, our results show that testing commercial products and optimizing protocols is very important and contribute for a better quality of products and services.Faculdade de Ciências e TecnologiaSilva, ZéliaVideira, PaulaRUNMarques, Graça Susete Costa de Carvalho2014-01-14T11:34:03Z20142014-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/11009enginfo: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:RCAAP2024-03-11T03:45:15Zoai:run.unl.pt:10362/11009Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:19:58.012704Repositório 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 Establishing a cell biology platform: isolation and preservation of human blood products
title Establishing a cell biology platform: isolation and preservation of human blood products
spellingShingle Establishing a cell biology platform: isolation and preservation of human blood products
Marques, Graça Susete Costa de Carvalho
Sera
Cell isolation
Cryopreservation
Cell culture
PBMCs (peripheral blood mononuclear cells)
title_short Establishing a cell biology platform: isolation and preservation of human blood products
title_full Establishing a cell biology platform: isolation and preservation of human blood products
title_fullStr Establishing a cell biology platform: isolation and preservation of human blood products
title_full_unstemmed Establishing a cell biology platform: isolation and preservation of human blood products
title_sort Establishing a cell biology platform: isolation and preservation of human blood products
author Marques, Graça Susete Costa de Carvalho
author_facet Marques, Graça Susete Costa de Carvalho
author_role author
dc.contributor.none.fl_str_mv Silva, Zélia
Videira, Paula
RUN
dc.contributor.author.fl_str_mv Marques, Graça Susete Costa de Carvalho
dc.subject.por.fl_str_mv Sera
Cell isolation
Cryopreservation
Cell culture
PBMCs (peripheral blood mononuclear cells)
topic Sera
Cell isolation
Cryopreservation
Cell culture
PBMCs (peripheral blood mononuclear cells)
description Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina
publishDate 2014
dc.date.none.fl_str_mv 2014-01-14T11:34:03Z
2014
2014-01-01T00:00:00Z
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/10362/11009
url http://hdl.handle.net/10362/11009
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
_version_ 1799137842088116224

Registros relacionados