Comparison of media components and reagents in cultivation and genetic editing of human cells A549

The work presents a comparative analysis of imported and Russian components of media and reagents in the cultivation and genetic editing of human lung adenocarcinoma A549 cells. The study used fetal bovine serum (FBS) from Gibco (USA), Intl Kang (China) and Technozerg (Russia), transfection reagents Lipofectamine3000 (USA) and GenJect39 (Russia), as well as OptiMem (USA) and FectoMEM (Russia) media. Cells cultured on all types of FBS retained high proliferative activity and survival, but the type of FBS had a significant effect on cell morphology. In particular, a fibroblast-like morphology was observed on the Technozerg medium. Transfection efficiency was assessed using the empty pCMV-EGFP expression vector and the CRISPR/Cas9-mediated human lambda DNA polymerase gene editing vector PX458-POLL. OptiMem and FectoMEM media maintained high transfection efficiency with either vector. GenJect39 was as efficient as Lipofectamine3000 for pCMV-EGFP transfection, but GenJect39 was significantly less efficient than Lipofectamine3000 for PX458-POLL genome editing. In addition, the choice of serum was shown to have a significant impact on the ability of cells to form colonies after PX458-POLL transfection. These results indicate that the choice of transfection reagent and FBS play an important role in successful experiment.