Investigation of the efficiency of CHO DP-12 cell transfection in media enriched with flaxseed protein hydrolysates

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.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