magistrsko delo
Kaja Marc (Author), Urban Švajger (Reviewer), Mojca Narat (Mentor), Mojca Narat (Thesis defence commission member), Sabina Vatovec (Thesis defence commission member), Urban Švajger (Thesis defence commission member), Polona Jamnik (Thesis defence commission member), Sabina Vatovec (Co-mentor)

Abstract

Sesalske celične linije se pogosto uporabljajo za industrijsko proizvodnjo biofarmacevtskih učinkovin. Celične banke predstavljajo zalogo celične kulture na dolgi rok, pri čemer z zamrzovanjem celicam na nek način ustavimo biološki čas. Iz celične banke lahko kadarkoli vzpostavimo nov bioproces. Da lahko zagotavljamo stabilnost zamrznjene celične kulture, je potrebno celice ustrezno pripraviti na zamrzovanje, saj ob zamrzovanju pride do tvorbe ledu oziroma kristalov zunaj in znotraj celice ter dehidracije, kar vodi v celične poškodbe. V ta namen dodajamo hitro penetrirajoč krioprotektant DMSO, ki ščiti celice pred poškodbami, vendar pa je hkrati, če je v previsoki koncentraciji ali če so mu celice predolgo izpostavljene, zanje toksičen. V magistrskem delu smo testirali, ali bi z nižjo koncentracijo DMSO lahko zmanjšali njegovo toksičnost na celice in tako povečali njihovo preživelost. Ugotovili smo, da bi bilo mogoče ob nižji uporabljeni koncentraciji DMSO med 4,5 in 7 % zamrzniti tudi višje koncentracije celic do 100×106 celic/ml. Visoko preživelost celic smo uspeli obdržati s pripravo zamrzovalne suspenzije v hladnih pogojih, s čimer smo podaljšali čas, ko so celice DMSO lahko izpostavljene pred zamrzovanjem. Pri tem se je močno zmanjšala tudi variabilnost med paralelkami.

Keywords

celične banke;zamrzovanje;sesalske celice;optimizacija;CHO;krioprotektanti;DMSO;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [K. Marc]
UDC: 606:62:57.086.13(043.2)
COBISS: 8972409 Link will open in a new window
Views: 1201
Downloads: 421
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Other data

Secondary language: English
Secondary title: Optimization of cryopreservation of chinese hamster ovary cells
Secondary abstract: Mammalian cell lines are often used for the industrial production of biopharmaceuticals. Cell banks represent the stock of cell culture in the long run, with the freezing of cells in some way stopping the biological time. From a cell bank we can establish new bioprocesses at any time. In order to ensure the stability of the frozen cell culture, the cells should be properly prepared for freezing, as when ice freezes, ice and crystals form outside and inside the cell and dehydrate, leading to cell damage. To this end, we add fast-penetrating cryoprotectant DMSO, which protects the cells against damage, but at the same time, if it is too high or if cells are too exposed to it, it is toxic for them. In the master's thesis, we tested whether a lower concentration of DMSO could reduce its cell toxicity and thus increase their survival. It was found that at a lower DMSO concentration used between 4.5 and 7 %, higher cell concentrations of up to 100×106 cells/ml could be frozen. We managed to maintain high survival of the cells by preparing a freeze-up suspension in cold conditions, thus extending the time when the DMSO cells could be exposed to freezing. Variability among parallels also greatly decreased.
Secondary keywords: cell banks;cryopreservation;mammalian;optimization;cryoprotectants;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Pages: XII, 83 f., [10] f. pril.
ID: 10919123