Monitoring cell cultures in real time in a biochip

• Autor(a) principal: Serrão, Ricardo Filipe Soares
• 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
• topic: Microfluidics; Cell Culture; Cell Viability; Adenylate Kinase; Photomultiplier; Immunoassay; Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias

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.