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Optimizacija zamrzovanja celic ovarija kitajskega hrčka
Dorn, Romana (Author), Narat, Mojca (Mentor) More about this mentor... This link opens in a new window, Vatovec, Sabina (Co-mentor)

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Abstract
Celice ovarija kitajskega hrčka se v farmacevtski industriji pogosto uporabljajo za proizvodnjo biofarmacevtskih učinkovin. Del procesa je proizvodnja celičnih bank, kjer se celice pripravi na zamrzovanje in dolgotrajno shranjevanje na ultra nizki temperaturi. Tako shranjene celice so po odtaljevanju v dobrem stanju in ohranjajo lastnosti celične linije. Zaradi tvorbe ledu med procesom zamrzovanja ali odtaljevanja lahko pride do poškodb celic, kar lahko vodi v nižjo viabilnost celic po odtaljevanju. Nastanek poškodb lahko preprečimo z dodatkom krioprotektantov, ki pa so lahko za celice po določenem času izpostavljenosti in koncentraciji toksični. V magistrski nalogi smo testirali različne koncentracije krioprotektanta DMSO, čase izpostavljenosti krioprotektantu pred zamrzovanjem ter temperaturo priprave zamrzovalne suspenzije z namenom optimizacije zamrzovanja. Testirali smo tudi, ali so ti dejavniki vplivali na titer produkta ob koncu bioprocesa z dohranjevanjem. Zaradi izpostavljenosti različnim temperaturam med rokovanjem s kriovialami smo testirali tudi učinke različnih temperatur na nekaterih celičnih linijah, shranjenih pri ultra nizki temperaturi. Ugotovili smo, da celice dosežejo višjo viabilnost, če pripravimo zamrzovalno suspenzijo pri hladnih pogojih. Nizka temperatura omogoča tudi daljši čas izpostavljenost celic krioprotektantu. Učinek različnih temperatur med rokovanjem s kriovialami na dolgotrajno shranjene celice je specifičen za celično linijo.

Language:Slovenian
Keywords:celične banke, zamrzovanje, sesalske celice, optimizacija, CHO, krioprotektanti, DMSO
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2020
Publisher:[R. Dorn]
UDC:606:62:57.086.13(043.2)
COBISS.SI-ID:32335619 This link opens in a new window
Views:309
Downloads:73
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Secondary language

Language:English
Title:Optimization of cryopreservation of chinese hamster ovary cells
Abstract:
Chinese hamster ovary cells are often used in a pharmaceutical industry for the production of biopharmaceuticals. Part of the process is cell banking, where the cells are prepared for freezing and long-term storage to maintain the cell line in original form and condition. Ice formation during freezing is causing injuries to the cells, which can result in low viability after thawing. As a prevention we add cryoprotectants, which can be toxic for cells, depending on the concentrations and exposure time. In the work presented, we aimed to optimize the process of cell bank preparation by testing different concentrations of cryoprotectant DMSO, its exposure time of cryoprotectant before freezing and temperature of preparation. We also tested whether these factors affected titer of product at the end of fed batch bioprocess. Because of exposure to different temperatures during handling with cryovials, we also tested the effect of different temperatures on cells stored at ultra-low temperature on several cell lines. We conclude that low temperature during cell bank preparation can result in higher viability and allows longer exposure time to cryoprotectant. Effect of different temperatures during handling of cryovials after long term storage on viabilities after thawing is cell line dependent.

Keywords:cell banks, cryopreservation, mammalian cells, optimization, CHO, cryoprotectants, DMSO

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