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.
Highlights
Cell-based therapy provides a cure to repair tissue damage and several degenerative diseases
The proliferation of human mesenchymal stem cells’ (hMSCs) with osteogenic medium under different culture environments was determined based on total DNA quantification (Figure 1a)
This study showed that hMSCs had a higher expansion rate than the static culture, even when the osteogenic medium was provided in these two dynamic culture systems (Figure 1, Table 1)
Summary
Cell-based therapy provides a cure to repair tissue damage and several degenerative diseases. It is known that terminally differentiated cells, such as chondrocytes and nucleus pulposus cells, may de-differentiate and lose their functional phenotype during in vitro cell culture expansion [2,3]. Rapid aging, phenotype change, and stemness loss are noticed in bone marrow-derived mesenchymal stem cells (MSCs) after in vitro expansion [4]. The culture conditions during monolayer cell expansion are known to affect the self-renewal, multipotency, and lineage-specific differentiation potentials of the adipose-derived stem cells [5]. Culture methods to preserve the differential potency of stem cells or to maintain a functional phenotype of terminally differentiated cells should be optimized for cell-based therapy
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