The Effects of Static and Dynamic Culture Systems on Cell Proliferation and Conditioned Media of Umbilical Cord-Derived Mesenchymal Stem Cells

Preclinical and clinical studies have demonstrated the therapeutic effects of umbilical cord-derived mesenchymal stem cells (UC-MSCs) and secretome to cure various degenerative diseases. Thus, the mass-scale production of MSCs is necessary to ensure their availability and costeffectiveness. In the current study, we evaluated the effect of dynamic 3D and static 2D culture systems on cell proliferation and conditioned media of UC-MSCs. The lysate of concentrated thrombocyte was used to substitute animal-derived serum in the culture media. From two experimental sets with different UC and lysates of concentrated thrombocyte donors, it was found that the shortest PDTs for experimental set 1 were 12.3 h (2D culture) and 14.8 h (3D culture), whereas in experimental set 2, they were 17.7 h (2D culture) and 16.9 h (3D culture). Microscopic observation confirmed the formation of cell aggregates in the 3D system, particularly during the exponential phase. SDS-PAGE analysis revealed similar protein profiles of conditioned media from both culture systems. An anti-inflammatory cytokine, namely tumor necrosis factor beta (TGF-β), was analyzed using ELISA to evaluate the effect of culturing methods on TGF-β release. Interestingly, the relative TGF-β contents in the 2D culture were stagnant throughout the incubation times, whereas a higher accumulation of TGF-β was detected in the 3D culture, which was most likely caused by shear stress. Our study confirmed that a dynamic culture system with a microcarriersupported bioreactor is a promising approach to scaling up MSC and secretome productions.