Rast i produktivnost CHO DP-12 stanica u kemijski definiranim hranjivim medijima

undergraduate thesis

• Title (english): Growth and Productivity of CHO DP-12 Cells in Chemically Defined Media
• Author: Ana Jelić
• Mentor: Igor Slivac (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: 2020-09-01, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Kulture CHO (eng. Chinese Hamster Ovary) stanica za proizvodnju rekombinantnih proteina zahtijevaju hranjive medije koji potiču rast stanica i sintezu proizvoda. Takvi mediji moraju osigurati visoku vijabilnost i koncentraciju stanica, a ujedno stimulirati sintezu i izlučivanje proizvoda izvan stanice. U ovom radu ispitivan je utjecaj kemijski definiranih medija Dynamis, CD CHO, EX-CELL i PowerCHO na rast i produktivnost stanične linije CHO DP-12 u suspenzijskim kulturama uzgajanim u tikvicama (eng. shake-flask). U svrhu mjerenja koncentracije i vijabilnosti stanica u kulturi korišten je hemocitometar i bojilo tripan-plavo. Koncentracije supstrata (glukoze) i metabolita (laktata) u mediju mjerene su spektrofotometrijski. Procjena produktivnosti staničnih kultura analizirana je prema prisutnosti rekombinantnog humanog monoklonskog protutijela anti-IL8 detektiranog elektroforezom (SDS-PAGE) u medijima u kojima su uzgajane stanice. Rezultati pokazuju da se najbolji rast i najveća produktivnost postižu u mediju Dynamis, dok u medijima CD CHO i EX-CELL nije postignut zadovoljavajući rast stanica što znači da je za uzgoj u njima potrebna stupnjevita prilagodba.

Abstract (english)

The culture of CHO cells (Chinese Hamster Ovary) requires media that support growth and recombinant protein production. Such media must support high viable cell densities while ensuring synthesis of desired products. The aim of this study was to examine the effect of chemically defined media: Dynamis, CD CHO, EX-CELL and PowerCHO, on growth and productivity of CHO DP-12 cell line in suspension culture in shake-flasks. Cell concentration and viability were determined with hemocytometer and trypan blue. Spectrophotometry was used for measuring glucose and lactate concentrations in media. Production of recombinant human anti-IL8 monoclonal antibody was evaluated using SDS-PAGE. The study showed that the best growth and highest productivity were achieved in Dynamis medium. CD CHO and EX-CELL media did not result in satisfactory cell growth indicating that gradual adaptation to these media is required.

• Keywords: CHO DP-12 stanična linija; kemijski definirani hranjivi medij; monoklonsko protutijelo; rast stanica
• Keywords (english): CHO DP-12 cell line; cell growth; chemically defined media; monoclonal antibody
• Language: croatian
• URN:NBN: urn:nbn:hr:159:258410
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-11-03 13:08:26