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

Abstract

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 cost-effectiveness. 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-?), was analyzed using ELISA to evaluate the effect of culturing methods on TGF-? release. Interestingly, the relative TGF-? contents in the 2D culture were stagnant throughout the incubation times, whereas a higher accumulation of TGF-? was detected in the 3D culture, which was most likely caused by shear stress. Our study confirmed that a dynamic culture system with a microcarrier-supported bioreactor is a promising approach to scaling up MSC and secretome productions.

Keywords

• bioreactor
• dynamic culture
• mesenchymal stem cells
• proliferation
• secretome