The Effects of Different Dynamic Culture Systems on Cell Proliferation and Osteogenic Differentiation in Human Mesenchymal Stem Cells

Abstract

The culture environment plays an important role for stem cells’ cultivation. Static or dynamic culture preserve differential potentials to affect human mesenchymal stem cells’ (hMSCs) proliferation and differentiation. In this study, hMSCs were seeded on fiber disks and cultured in a bidirectional-flow bioreactor or spinner-flask bioreactor with a supplement of osteogenic medium. The hMSCs’ proliferation, osteogenic differentiation, and extracellular matrix deposition of mineralization were demonstrated. The results showed that the spinner flask improved cell viability at the first two weeks while the bidirectional-flow reactor increased the cell proliferation of hMSCs through the four-week culture period. Despite the flow reactor having a higher cell number, a lower lactose/glucose ratio was noted, revealing that the bidirectional-flow bioreactor provides better oxygen accessibility to the cultured cells/disk construct. The changes of calcium ions in the medium, the depositions of Ca2+ in the cells/disk constructs, and alkaline phosphate/osteocalcin activities showed the static culture of hMSCs caused cells to mineralize faster than the other two bioreactors but without cell proliferation. Otherwise, cells were distributed uniformly with abundant extracellular matrix productions using the flow reactor. This reveals that the static and dynamic cultivations regulated the osteogenic process differently in hMSCs. The bidirectional-flow bioreactor can be used in the mass production and cultivation of hMSCs for applications in bone regenerative medicine.

Induction of osteogenic differentiation of bone marrow stromal cells on 3D polyester-based scaffolds solely by subphysiological fluidic stimulation in a laminar flow bioreactor

“…The upregulation was observed without the presence of osteogenic supplements and (b) putative MSC markers, CD73 and CD90, were significantly downregulated in the FL-L and FL-H. affect cell growth. 13,50 Indeed, air bubbles are readily generated in perfusion bioreactors used for bone tissue engineering due to continuous flow, serum proteins acting as a surfactant and the microporous structure and hydrophobic nature of 3D scaffolds. [51][52][53][54] Therefore, Henry's law was applied to supress bubble formation completely.…”

“…These impede fluid flow, and bubbles entrapped within the scaffolds may adversely affect cell growth. 13 , 50 Indeed, air bubbles are readily generated in perfusion bioreactors used for bone tissue engineering due to continuous flow, serum proteins acting as a surfactant and the microporous structure and hydrophobic nature of 3D scaffolds. 51 – 54 Therefore, Henry’s law was applied to supress bubble formation completely.…”

“…However, most of these studies were conducted in the presence of either chemical supplements or osteoinductive biomaterials such as decellularised matrix constructs, ECM-coated or hydroxyapatite-laden scaffolds. 13,[21][22][23][24][25] The lack of concrete evidence supporting fluid flow-induced osteogenesis in 3D scaffolds may be also attributed to the complexity of 3D dynamic culture systems. A relatively complex bioreactor set-up is required to establish stable culture conditions such as systems for environmental monitoring and control.…”

“… 11 , 12 Laminar flow bioreactors facilitate precise control of fluid pattern through engineered constructs, which is considered to be a less damaging and more predictable procedure. 13 Moreover, laminar flow bioreactors provide uniform gas and nutrient supply within the construct with a relatively low shear rate. 6 Therefore, the use of laminar flow bioreactors seems advantageous for controlling the fate of progenitors by fluidic stimuli.…”

Clarifying the Tooth-Derived Stem Cells Behavior in a 3D Biomimetic Scaffold for Bone Tissue Engineering Applications

“…Dynamic culture has previously shown to positively affect human MSCs proliferation, differentiation, and ECM production when compared to static 3D cultures ( Woloszyk et al, 2014 ). A dynamic culture system should improve nutrients and oxygen diffusion, avoiding hypoxia-induced central necrosis in cultured tissue constructs ( Tsai et al, 2019 ). However, some bioreactors could promote hydrodynamic stress during cell culture resulting in shear stress, thus causing cell damage ( Tanzeglock et al, 2009 ).…”