Abstract
Osteocytes are mechanosensory commander cells to regulate bone remodeling throughout the lifespan. While the osteocytes are known as terminally differentiated cells derived from mesenchymal stem cells, the detailed mechanisms of osteocyte differentiation remain unclear. In this study, we fabricated 3D self-organized spheroids using human mesenchymal stem cells (MSCs). Under the osteogenesis induction medium, the spheroid culture model exerted the osteocyte-likeness within 2 days compared to a conventional 2D monolayer model. Moreover, we showed that an inhibition of actin polymerization in the spheroid further up-regulated the osteocyte gene expressions. Notably, we represented that the cell condensed condition acquired in the 3D spheroid culture model determined a differentiation fate of MSCs to osteocytes. Taken together, we suggest that our self-organized spheroid model can be utilized as a new in vitro model to represent the osteocyte and to recapitulate an in vitro ossification process.
Highlights
Osteocytes are mechanosensory commander cells to regulate bone remodeling throughout the lifespan
We showed that the osteocyte differentiation of pre-osteoblast cells was facilitated in the 3D scaffold-free culture system[14,15,16]
We revealed that cell condensed condition is significant for the pre-osteoblast cells to undergo the osteocyte differentiation, it is still unclear whether or not the osteocyte differentiation can be further evoked from the mesenchymal stem cells to recapitulate the in vivo bone development process
Summary
Osteocytes are mechanosensory commander cells to regulate bone remodeling throughout the lifespan. After the osteoblasts produce the bone matrices such as collagen type I and alkaline phosphatase during the bone remodeling p rocess[6,7], it is known that they encounter one of the fates; (1) to become bone-lining cells, (2) to become osteocytes embedded in the bone, and (3) to undertake apoptosis[2,8] For both osteoblast differentiation and bone mineralization studies, many researchers have established a gold standard protocol to induce in vitro osteogenesis using chemical osteogenesis induction (OI) supplements such as ascorbic acid and β-glycerophosphate[9,10]. By using the MSC spheroid model, we attempted to elucidate the mechanism of the cell-fate decision of MSCs toward osteocytes and to recapitulate the in vitro ossification process derived from the mesenchymal condensation
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