Istraživanje učinkovitosti transfekcije CHO DP-12 stanica u mediju obogaćenom proteinskim hidrolizatima lana

master's thesis

• Title (english): Investigation of the efficiency of CHO DP-12 cell transfection in media enriched with flaxseed protein hydrolysates
• Author: Vlatka Kozarić
• Mentor: Igor Slivac (mentor)
• Committee member: Višnja Gaurina-Srček (predsjednik povjerenstva)
• Committee member: Andreja Leboš-Pavunc (član povjerenstva)
• Committee member: Teuta Murati (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); Zagreb
• Defense date and country: 2022-07-07, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Interdisciplinary Technical Sciences

Abstract

U posljednje vrijeme sve je veći interes za primjenom uljnih pogača sjemenki lana zbog njihovog visoko vrijednog sastava u kojem oko 20 % čine proteini. Kako je već ranije utvrđeno da proteinski hidrolizati lana potiču rast CHO DP-12 stanica, u ovom radu se otišlo korak dalje te je ispitan njihov utjecaj na uspješnost transfekcije CHO DP-12 stanica. Transfekcija se provodila i na adherentnim i na suspenzijskim stanicama. Korištena su dva transfekcijska sredstva, FuGene 6 i Turbofect te nekoliko različitih hranjivih medija (PowerCHO, DMEM, ExCell, CD CHO, FidCHO). Transfekcija je zabilježena samo u DMEM mediju uz FuGene 6 transfekcijsko sredstvo, bez dodatka hidrolizata lana. Ispitivanjem različitih omjera mase plazmida (μg) i transfekcijskog sredstva (mL), ustanovljeno je da najveću učinkovitost transfekcije daje omjer 1:6. Iako je utvrđeno da proteinski hidrolizati lana imaju negativan utjecaj na transfekciju CHO DP-12 stanica, dobiveni rezultati su dobra podloga za daljnja istraživanja u cilju pronalaska optimalnih uvjeta transfekcije.

Abstract (english)

The application of flaxseed oil cake attracts great interest due to its highly valuable composition and protein content of about 20 %. As it has been already known that flax protein hydrolysates stimulate the growth of CHO DP-12 cells, this work went a step further and their impact on the efficiency of CHO DP-12 cells transfection has been examined. Transfection was performed on both adherent and suspension cells. Two transfection reagents were used, FuGene 6 and Turbofect, and several different nutrient media (PowerCHO, DMEM, ExCell, CD CHO, FidCHO). Transfection was recorded only in DMEM medium with FuGene 6 transfection reagent, without adding flax hydrolyzates. Examination of different ratios of plasmid mass (μg) and transfection reagent (mL), revealed that the ratio of 1:6 gives the highest transfection efficiency. Although it has been determined that flax protein hydrolysates have a negative effect on CHO DP-12 cell transfection, the obtained results are good foundation for further research to find optimal transfection conditions.

• Keywords: transfekcija; proteinski hidrolizati; CHO DP-12 stanična linija; lan; hranjivi medij
• Keywords (english): transfection; protein hydrolysates; CHO DP-12 cell line; flaxseed; cell culture medium
• Language: croatian
• URN:NBN: urn:nbn:hr:159:003455
• 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: Open access
• Created on: 2023-01-19 13:19:35