Primjena mikronosača Cytodex 1 u proizvodnji biomase Chinese Hamster Ovary (CHO-K1) stanica

master's thesis

• Title (english): Application of cytodex 1 microcarriers in production of Chinese Hamster Ovary (CHO-K1) cell biomass
• Author: Marija Šiprak
• Mentor: Zlatko Kniewald (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: 2007-10-10, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

U ovom radu ispitane su mogućnosti uporabe Cytodex 1 mikronosača u proizvodnji biomase CHO-K1 stanica u spinner bioreaktoru, te je uspoređena dinamike rasta CHO-K1 stanica na novim i već jednom uporabljenim Cytodex 1 mikronosačima. Broj stanica je uspoređen brojanjem uz dodatak boje tripan-plavo i kristal-violet. Brojanjem CHO-K1 stanica uz dodatak boje kristal-violet utvrđeno je da stvaranje monosloja na uporabljenim Cytodex 1 mikronosačima bez dodatka svježeg inokuluma nije moguće. Praćenjem rasta CHO-K1 stanica uzgajanih u spinner bioreaktoru kroz 9 dana na novim i uporabljenim Cytodex 1 mikronosačima, kod jednakih uvjeta uzgoja, nije primjećena značajna razlika u kinetici kojom rastu niti u prihvaćanju na mikronosače i metabolizmu. U spinner bioreaktoru nacijepljenom s 4,2 x 107 CHO-K1 stanica i 0,3 g novih Cytodex 1 mikronosača, tijekom 9 dana uzgoja, postignut je prinos stanica od 2,1 x 108 stanica, uz specifičnu brzinu rasta od 0,019 h-1. U spinner bioreaktoru nacijepljenom s 4,2 x 107 CHO-K1 stanica i 0,3 g uporabljenih Cytodex 1 mikronosača, tijekom 9 dana uzgoja, postignut je prinos CHO-K1 stanica od 1,7 x 108 stanica, uz specifičnu brzinu rasta od 0,017 h-1. Koncentracija stanica određena brojanjem jezgri kristal-violetom veća je za 10-30 % od one određene metodom brojanja stanica tripan-plavim zbog prisustva binuklearnih stanica u kulturi.

Abstract (english)

This paper examines the possibility of using Cytodex 1 microcarriers when cultivating CHO-K1 cells in a spinner flask, and compares the growth dynamics of CHO-K1 cells on new and re-used Cytodex 1 microcarriers. The number of cells was determined by adding tripan-blue and crystal-violet dyes. By counting the cells stained with crystal-violet dye, it was determined that the origination of a monolayer on Cytodex 1 microcarriers without adding fresh inoculum is not possible. When monitoring the growth of CHO-K1 cells during 9 days, cultured in a spinner flask on new and re-used Cytodex 1 mycrocarriers, and in the same growth conditions, it was determined that there is not a significant difference in their growth kinetics, cell attachment and metabolism. In the spinner flask inoculated with 4.2 x 107 CHO-K1 cells and 0.3 g of new Cytodex 1 microcarriers cultivated during 9 days, a cell yield of 2.1 x 108 cells was reached with a specific growth rate of 0.019 h-1. In the spinner flask inoculated with 4.2 x 107 CHO-K1 cells and 0.3 g of re-used Cytodex 1 microcarriers cultivated during 9 days, a cell yield of 1.7 x 108 cells was reached with a specific growth rate of 0.017 h-1. Cell concentration determined by nuclear counts through crystal-violet staining is higher by 10-30 percent than the one determined by the tripan-blue cell counting method because binucleated cells are present in the culture.

• Keywords: Cytidex 1 mikronosači; CHO-K1 stanice; spinner bioreaktor; tripan-plavo; kristal-violet
• Keywords (english): Cytodex 1 microcarriers; CHO-K1 cells; spinner flask; tripan-blue dye; crystal-violet dye
• Language: croatian
• URN:NBN: urn:nbn:hr:159:165363
• Study programme: Title: Biotechnology; specializations in: Biochemical-Microbiology; Course: Biochemical-Microbiology; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki (diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-09-05 13:41:03