Abstract
Objectives
To investigate the behaviors of aggregates of human mesenchymal stem cells (hMSCs) on chondrogenesis and chondrocyte hypertrophy using spatiotemporal expression patterns of chondrogenic (type II collagen) and hypertrophic (type X collagen) markers during chondrogenesis.
Results
hMSCs were cultured on either a polystyrene surface or polyamidoamine dendrimer surface with a fifth generation (G5) dendron structure in chondrogenic medium and growth medium. At day 7, cell aggregates without stress fibers formed on the G5 surface and triggered differentiation of hMSCs toward the chondrogenic fate, as indicated by type II collagen being observed while type X collagen was undetectable. In contrast, immunostaining of hMSCs cultured on polystyrene, which exhibited abundant stress fibers and did not form aggregates, revealed no evidence of either type II and or type X collagen. At day 21, the morphological changes of the cell aggregates formed on the G5 surface were suppressed as a result of stress fiber formation. Type II collagen was observed throughout the aggregates whereas type X collagen was detected only at the basal side of the aggregates. Change of cell aggregate behaviors derived from G5 surface alone regulated chondrogenesis and hypotrophy, and this was enhanced by chondrogenic medium.
Conclusions
Incubation of hMSCs affects the expression of type II and X collagens via effects on cell aggregate behavior and stress fiber formation.
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Acknowledgements
The authors gratefully acknowledge financial support from the Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (Grant No. PHD/0016/2555). This work was also supported by the project “Development of cell manufacturing and processing system for industrialization of regenerative medicine” (No. P14006) commissioned by the Japan Agency for Medical Research and Development (AMED).
Supporting information
Supplementary Movie 1—Behavior of hMSC aggregates on G5 and polystyrene surfaces in growth medium and chondrogenic medium. The analysis was performed at an early phase of culture from day 6 to 7 and from day 20 to 21.
Supplementary Movie 2—Fluorescence images of type II collagen (green), nuclei (blue), and F-actin (red) in hMSCs cultured on G5 and polystyrene surfaces in growth medium at day 7 and 21.
Supplementary Movie 3—Fluorescence images of type X collagen (green), nuclei (blue), and F-actin (red) in hMSCs cultured on G5 and polystyrene surfaces in growth medium at day 7 and 21.
Supplementary Movie 4—Fluorescence images of type II collagen (green), nuclei (blue), and F-actin (red) in hMSCs cultured on G5 and polystyrene surfaces in chondrogenic medium at day 7 and 21.
Supplementary Movie 5—Fluorescence images of type X collagen (green), nuclei (blue), and F-actin (red) in hMSCs cultured on G5 and polystyrene surfaces in chondrogenic medium at day 7 and 21.
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Wongin, S., Ogawa, Y., Kim, MH. et al. Chondrogenesis and hypertrophy in response to aggregate behaviors of human mesenchymal stem cells on a dendrimer-immobilized surface. Biotechnol Lett 39, 1253–1261 (2017). https://doi.org/10.1007/s10529-017-2339-9
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DOI: https://doi.org/10.1007/s10529-017-2339-9