Development of Serum-Free Medium and Suspension Culture for HEK293 Cells Using Systematic Optimization Strategies

Cell culture plays a pivotal role in the biomedical industry, with a growing shift from traditional serum-containing adherent systems to serum-free suspension culture platforms. This transition underscores the importance of developing robust cell suspension adaptation protocols and serum-free culture media to meet industrial demands. In this study, we employed a combination of cell suspension acclimation, differential screening of culture media, and factorial design methodology to rapidly adapt adherent human embryonic kidney (HEK293) cells to suspension culture. Additionally, we developed a chemically defined medium optimized for these cells within a short timeframe. By systematically adjusting the concentrations of key medium components and integrating an optimized adaptation protocol, we successfully achieved efficient suspension culture of HEK293 cells. Notably, this approach significantly reduced cell aggregation and enhanced cell growth density, thereby advancing the potential for industrial-scale applications.