Prilagodba CCO stanične linije na rast u mediju bez dodatka seruma

master's thesis

• Title (english): Adaptation of CCO cell line growth in serum-free medium
• Author: Bogdanka Dukić
• Mentor: Višnja Gaurina Srček (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2012-09-24, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Proliferacija životinjskih stanica u kulturi pod kontrolom je vanjskih faktora rasta najčešće sadržanih u serumu životinjskog podrijetla. Korištenje životinjskog seruma se prilikom uzgoja nastoji svesti na najmanju moguću mjeru, radi nedovoljnog poznavanja sastava, nepoznatih mehanizama utjecaja na rast, interferencije s drugim spojevima u mediju, i sl. Kako bi se smanjilo njegovo korištenje, proizvode se mediji bez seruma koji osiguravaju zadovoljavajući rast stanica. Stanice se za uzgoj u takvom mediju moraju prilagoditi. Svrha ovog rada bio je pokušaj prilagodbe CCO stanične linije na rast u mediju bez dodatka seruma. CCO stanična linija spada u adherentni tip stanica koje su ovisne o površini za rast. Kako bi se poboljšao prinos stanica, osim u Multiwell pločama, proveden je i uzgoj na Cytodex 1 mikronosačima u Cell-spin bioreaktorima. Stanice su uzgajane u Multiwell pločama u mediju s različitim koncentracijama seruma postupnom prilagodbom od početnih 10 % do 0 % seruma. Prilagodba je bila uspješna, budući da je pri svakoj koncentraciji seruma utvrđen rast stanica, pri čemu je u mediju bez seruma utvrđen prinos stanica od 2 x105 st/mL. U Cell-spin bioreaktorima stanice su uzgajane na Cytodex 1 mikronosačima u dva različita medija, pri čemu je jedan bio DMEM medij s 5 % seruma koji je preporučen za uzgoj CCO stanica od strane proizvođača, a drugi UC medij sa smanjenom koncentracijom seruma od 2 %. Stanice su uspješno prilagođene na uzgoj u mediju sa smanjenom koncentracijom seruma, pri čemu je zabilježen prinos stanica od 17x105 st/mL.

Abstract (english)

Proliferation of animal cells in culture is under control of external growth factors usually present in animal serum. During cultivation it is desirable to minimalize serum usage because of unknown composition, mechanisms which can influence on growth, interfere with other substances present in media, etc. In order to reduce serum usage, serum-free media are produced to provide satisfying cell growth. The cells which grow in that sort of media must be adapted for that cultivation. The aim of this thesis was the attempt of adaptation of CCO cell line to cultivation in media without serum. CCO cell line cells are adherent which means that they can not grow without surface on which they attach. In order to improve cell yield, cells were grown not only on Multiwell plates, but they were cultivated also on Cytodex 1 microcarriers in Cell-spin bioreactors. On Multiwell plates cells were grown in media with different serum concentrations, and were gradually adapted to medium without serum, starting with medium containing 10 % of serum, until medium with 0 % of serum. Adaptation was successful because at every serum concentration cell growth was established, whereby in media without serum cell yield was 2x105 cells/mL. In Cell-spin bioreactors cells have been cultivated on Cytodex 1 microcarriers in two different media, whereby one has been DMEM media with 5 % of serum which is recomended for CCO cells cultivation by the producer, and the other one was UC media with reduced serum concentration of 2 %. The cells were successfully adapted to cultivation in media with reduced serum concentration, whereby gained cell yield was 17x105 cells/mL.

• Keywords: CCO stanična linija; Cell-spin bioreaktor; Cytodex 1 mikronosači; serum
• Keywords (english): CCO cell line; Cell-spin bioreactor; Cytodex 1 microcarriers; serum
• Language: croatian
• URN:NBN: urn:nbn:hr:159:820418
• Study programme: Title: Bioprocess Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-09 09:29:26