Publicação
In silico culture media costumization for Human Embrionic Kidney 293 Cells
| datacite.subject.fos | Engenharia e Tecnologia::Engenharia Química | pt_PT |
| dc.contributor.advisor | Oliveira, Rui | |
| dc.contributor.advisor | Costa, Rafael | |
| dc.contributor.author | Santos, Cíntia Dione Carvalho dos | |
| dc.date.accessioned | 2020-07-02T14:08:43Z | |
| dc.date.available | 2022-04-02T00:30:30Z | |
| dc.date.issued | 2020-06-03 | |
| dc.date.submitted | 2020 | |
| dc.description.abstract | Human Embryonic Kidney (HEK-293) cells have gained substantial interest in cell biology research and biotechnology. Still, there are not many studies in the literature on the specificities of the nutritional requirements and culture media composition to grow HEK-293 cells in comparison to other mammalian cells. As the development of a custom culture media for a specific cell line is, among many factors, dependent on the complexity of cellular physiology, a better understanding of the cellular metabolism can help to predict the response on a particular culture medium composition. With the goal to determine the culture medium composition on cellular expansion of HEK293 cells and develop a rational in silico design methodology of culture medium composition, a metabolic network for HEK-293 cells was established, based on the human genome scale model Recon-2. The metabolic model comprises 327 biochemical reactions and 341 metabolites.In order to validate the HEK-293 metabolic model, flux balance analysis (FBA) and flux variability analysis (FVA) methods were performed in MATLAB based on a constrained nonlinear program. The computed flux distributions and respective 95% confidence intervals were compared/validated with experimental data from the literature. The computed flux distributions for the first and second metabolic phases were highly concordant with literature values (explained variance > 85%). In a few cases, significative differences were observed. These exceptions were analyzed in detail. An hypothetical minimum consumption medium scenario was computed, where substrates uptake are minimized concomitantly to the minimization of toxic by-products formation. Results demonstrate that 100% of lactate and ammonia production can be ceased as a consequence of glucose and non-essential amino-acids (AA) intake reduction. However, reduction of byproduct formation did not necessarily result in an improved cell’s energetic efficiency, as ATP synthase did not increase with regards of lactate dehydrogenase reduction. | pt_PT |
| dc.identifier.uri | http://hdl.handle.net/10362/100218 | |
| dc.language.iso | eng | pt_PT |
| dc.subject | Culture Media Optimization | pt_PT |
| dc.subject | HEK-293 cells | pt_PT |
| dc.subject | Flux Balance Analysis | pt_PT |
| dc.subject | Flux Variability Analysis | pt_PT |
| dc.title | In silico culture media costumization for Human Embrionic Kidney 293 Cells | pt_PT |
| dc.type | master thesis | |
| dspace.entity.type | Publication | |
| rcaap.rights | openAccess | pt_PT |
| rcaap.type | masterThesis | pt_PT |
| thesis.degree.name | Mestre em Engenharia Química e Bioquímica | pt_PT |
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