University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal
Gewählte Master / Diploma Thesis:
Georg Hackl
(2021):
Maintenance Energy
Critical Reassessment and Determination in CHO.
Master / Diploma Thesis - Institut für Tierische Zelltechnologie und Systembiologie,
BOKU-Universität für Bodenkultur,
pp 53.
UB BOKU
obvsg
- Abstract:
- In the last century, there was a lot of research going into new isolated cells for industrial production. As Tjio and Puck discovered “Chinese hamster ovary cells” from the organism Cricetulus griseus in 1958, they predicted their great potential for industrial application. As expected, their use grew steadily to the major producing cell. Nowadays, many pharmaceuticals are produced by CHO cells. Since there are a few powerful tools to increase their production, they are used frequently in industry. Today, these producer cells are already well researched, but need further investigation to give deeper insight in their behaviour regarding various environmental conditions but also intrinsic factors. In collaboration of various universities, the genome and metabolism of CHO cells were investigated. This work led to a CHO-specific Genome-scale-metabolic-model (GSMM). Since a few factors are incorporated insufficiently, this model needs further improvement. This work gives valuable insight on the value of this model, which is necessary to be looked at further. In particular, this thesis concentrates on maintenance energy. All organisms have this not negligible factor of energy demand, which is needed to keep the basic cell functions running. Therefore, it reduces the available energy for the cell, that can be used for production. Therefore, CHO cells (CHO, glutamine synthetase negative, Herceptin producer) were cultivated continuously in CD-CHO medium, supplemented with 0.2% Anti-Clumping Agent and 8 mM L-Glutamine. Then, two dilution rates were carried out (D = 0.029 h^(-1) and 0.021 h^(-1)) and the concentration of several metabolites and the product (Ammonium, L-Glutamine, L-Glutamate, D-Glucose, L-Lactate, other Amino acids, Herceptin) were measured in steady state. Additionally, samples for RNA-sequencing were also taken. Then, these values can be used as input of the GSMM. The further evaluation of all the samples is currently in process.
- Beurteilende(r): Borth Nicole