Monitoring cell cultures in real time in a biochip
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/95872 |
Resumo: | Drug screening is a very important procedure in the approval of drugs for cancer treatment. This process is generally carried out using in vitro or in vivo models that aren’t very efficient due to the non-reproducibility of the cellular and/or tissue microenvironment and ethical issues due to the use of animal models. Additionally, drug approval is a process that could last 10 to 15 years, too much time when therapy is required with urgency. Microfluidic structures can address such issues, decreasing the time per assay, as well as decreasing the quantity of reagents used and the volume of waste generated, thus decreasing the costs. Also, due to the generation of concentration gradients inside a microfluidic device, it mimics the microenvironment characteristic of conventional cell culture. In this work, a reproducible cell culture of HCT-116, a human colon cancer cell line, is successfully grown inside a microfluidic device for a posterior exposure to anti-cancer drugs. The cell viability, detected through staining the DNA with fluorophores, is on average 90%. To monitor the cell death via exposure to drugs, a specific cell death biomarker, adenylate kinase (AK), is detected inside a microfluidic device using a photomultiplier and a fluorescence microscope in a chip-based immunoassay. AK concentrations near the concentrations of the enzyme released by dead cells were detected with the immunoassay by concentrating the AK in packed agarose beads inside de microfluidic structure. |
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Monitoring cell cultures in real time in a biochipMicrofluidicsCell CultureCell ViabilityAdenylate KinasePhotomultiplierImmunoassayDomínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e TecnologiasDrug screening is a very important procedure in the approval of drugs for cancer treatment. This process is generally carried out using in vitro or in vivo models that aren’t very efficient due to the non-reproducibility of the cellular and/or tissue microenvironment and ethical issues due to the use of animal models. Additionally, drug approval is a process that could last 10 to 15 years, too much time when therapy is required with urgency. Microfluidic structures can address such issues, decreasing the time per assay, as well as decreasing the quantity of reagents used and the volume of waste generated, thus decreasing the costs. Also, due to the generation of concentration gradients inside a microfluidic device, it mimics the microenvironment characteristic of conventional cell culture. In this work, a reproducible cell culture of HCT-116, a human colon cancer cell line, is successfully grown inside a microfluidic device for a posterior exposure to anti-cancer drugs. The cell viability, detected through staining the DNA with fluorophores, is on average 90%. To monitor the cell death via exposure to drugs, a specific cell death biomarker, adenylate kinase (AK), is detected inside a microfluidic device using a photomultiplier and a fluorescence microscope in a chip-based immunoassay. AK concentrations near the concentrations of the enzyme released by dead cells were detected with the immunoassay by concentrating the AK in packed agarose beads inside de microfluidic structure.Conde, PedroRUNSerrão, Ricardo Filipe Soares2020-04-08T13:46:13Z2019-1120192019-11-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/95872enginfo: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:RCAAP2024-03-11T04:43:49Zoai:run.unl.pt:10362/95872Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:38:28.171526Repositório 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 |
Monitoring cell cultures in real time in a biochip |
title |
Monitoring cell cultures in real time in a biochip |
spellingShingle |
Monitoring cell cultures in real time in a biochip Serrão, Ricardo Filipe Soares Microfluidics Cell Culture Cell Viability Adenylate Kinase Photomultiplier Immunoassay Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
title_short |
Monitoring cell cultures in real time in a biochip |
title_full |
Monitoring cell cultures in real time in a biochip |
title_fullStr |
Monitoring cell cultures in real time in a biochip |
title_full_unstemmed |
Monitoring cell cultures in real time in a biochip |
title_sort |
Monitoring cell cultures in real time in a biochip |
author |
Serrão, Ricardo Filipe Soares |
author_facet |
Serrão, Ricardo Filipe Soares |
author_role |
author |
dc.contributor.none.fl_str_mv |
Conde, Pedro RUN |
dc.contributor.author.fl_str_mv |
Serrão, Ricardo Filipe Soares |
dc.subject.por.fl_str_mv |
Microfluidics Cell Culture Cell Viability Adenylate Kinase Photomultiplier Immunoassay Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
topic |
Microfluidics Cell Culture Cell Viability Adenylate Kinase Photomultiplier Immunoassay Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
description |
Drug screening is a very important procedure in the approval of drugs for cancer treatment. This process is generally carried out using in vitro or in vivo models that aren’t very efficient due to the non-reproducibility of the cellular and/or tissue microenvironment and ethical issues due to the use of animal models. Additionally, drug approval is a process that could last 10 to 15 years, too much time when therapy is required with urgency. Microfluidic structures can address such issues, decreasing the time per assay, as well as decreasing the quantity of reagents used and the volume of waste generated, thus decreasing the costs. Also, due to the generation of concentration gradients inside a microfluidic device, it mimics the microenvironment characteristic of conventional cell culture. In this work, a reproducible cell culture of HCT-116, a human colon cancer cell line, is successfully grown inside a microfluidic device for a posterior exposure to anti-cancer drugs. The cell viability, detected through staining the DNA with fluorophores, is on average 90%. To monitor the cell death via exposure to drugs, a specific cell death biomarker, adenylate kinase (AK), is detected inside a microfluidic device using a photomultiplier and a fluorescence microscope in a chip-based immunoassay. AK concentrations near the concentrations of the enzyme released by dead cells were detected with the immunoassay by concentrating the AK in packed agarose beads inside de microfluidic structure. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-11 2019 2019-11-01T00:00:00Z 2020-04-08T13:46:13Z |
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/10362/95872 |
url |
http://hdl.handle.net/10362/95872 |
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.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 |
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1799138001081597952 |