Abstract
The goal of our research work is to establish mesenchymal osteoprogenitors derived from human jaw periosteum for tissue engineering applications in oral and maxillofacial surgery. For future autologous and/or allogeneic transplantations, some issues must be addressed. On the one hand, animal-free culture conditions have yet to be established. On the other hand, attempts should be undertaken to shorten the in vitro culturing process efficiently. The aim of the present study is to compare and analyze the phenotype of osteoprogenitors from the jaw periosteum under normal FCS-containing and animal-free culture conditions. Therefore, we analyzed the proliferation rates of MesenCult-XF medium (MC-) in comparison to DMEM-cultured JPCs. Whereas jaw periosteal cells (JPCs) show relatively slow proliferation rates and a fibroblastoid shape under DMEM culture conditions, MC-cultured JPCs diminished their cell size significantly and proliferated rapidly. By live-monitoring measurements of adhesion and proliferation, we made an interesting observation: whereas the proliferation of the MSCA-1+ subpopulation and the unseparated cell fraction were favored by the animal-free culture medium, the proliferation of the MSCA-1- subpopulation seemed to be repressed under these conditions. The alkaline phosphatase expression pattern showed similar results under both culture conditions. Comparison of the mineralization capacity revealed an earlier formation of calcium-phosphate precipitates under MC culture conditions; however, the mineralization capacity of the DMEM-cultured cells seemed to be higher. We conclude that the tested animal-free medium is suitable for the in vitro expansion and even for the specific selection of osteoprogenitor cells derived from the jaw periosteum.
Highlights
In previous studies, we characterized the phenotype, osteogenic potential and features of jaw periosteal cells (JPCs) grown in twoand three-dimensional culture conditions [1,2,3,4,5,6,7]
We focused the present study on the comparison of the JPC phenotype under FCS-containing and FCS-free culture conditions and analyzed in detail the proliferation and mineralization capacities of the cells, as well as their expression of common stem cell and osteogenesis-relevant markers
We detected an earlier formation of calcium phosphate precipitates in MC-XF-cultured JPCs at day 12 of osteogenesis
Summary
We characterized the phenotype, osteogenic potential and features of jaw periosteal cells (JPCs) grown in twoand three-dimensional culture conditions [1,2,3,4,5,6,7]. We determined that not all isolated jaw periosteal cells are able to mineralize in vitro. Due to this fact, the identification of specific markers for the osteogenic progenitors from the jaw periosteum is of paramount importance for the selection of these cells from the whole heterogeneous cell population. Screenings for cell surface markers revealed that two mesenchymal stem cell markers were significantly induced during the osteogenesis of jaw periosteal cells. The cell surface antigen mesenchymal stem cell antigen-1 (MSCA-1) was shown to be significantly more induced in mineralizing compared to non-mineralizing jaw periosteal cells, both in the undifferentiated and differentiated states. The shortening of the in vitro expansion procedure, as well as the use of serum-free culture conditions, should be achieved to ensure the success of future tissue engineering applications using cell-based constructs
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