Abstract
To achieve safer patient treatments, serum-free cell culture conditions have to be established for cell therapies. In previous studies, we demonstrated that serum-free culture favored the proliferation of MSCA-1+ osteoprogenitors derived from the jaw periosteum. In this study, the in vitro formation of bone-specific matrix by MSCA-1+ jaw periosteal cells (JPCs, 3 donors) was assessed and compared under serum-free and serum-containing media conditions using the marker-free Raman spectroscopy. Based on a standard fluorescence assay, JPCs from one patient were not able to mineralize under serum-containing culture conditions, whereas the other cells showed similar mineralization levels under both conditions. Raman spectra from mineralizing MSCA-1+ JPCs revealed higher levels of hydroxyapatite formation and higher mineral to matrix ratios under serum-free culture conditions. Higher carbonate to phosphate ratios and higher crystallinity in JPCs cultured under serum-containing conditions indicated immature bone formation. Due to reduced collagen production under serum-free conditions, we obtained significant differences in collagen maturity and proline to hydroxyproline ratios compared to serum-free conditions. We conclude that Raman spectroscopy is a useful tool for the assessment and noninvasive monitoring of in vitro mineralization of osteoprogenitor cells. Further studies should extend this knowledge and improve JPC mineralization by optimizing culture conditions.
Highlights
Jaw periosteum-derived osteoprogenitor cells (JPCs) represent an optimal stem cell source for bone tissue engineering applications in oral and maxillofacial surgeries
We showed previously that a subpopulation of JPCs expressing a high level of mesenchymal stem cell antigen-1 (MSCA-1) was shown to exhibit an increased osteogenic potential compared to the MSCA-1low cell fraction [1]
We described that JPCs show higher proliferation rates and a higher expression of the osteoprogenitor marker MSCA-1 after expansion under serum-free culture conditions [2]
Summary
Jaw periosteum-derived osteoprogenitor cells (JPCs) represent an optimal stem cell source for bone tissue engineering applications in oral and maxillofacial surgeries. Detailed characterization of JPCs, optimized serum-free culture, and differentiation conditions must be established to license these cells for patient studies. We showed previously that a subpopulation of JPCs expressing a high level of mesenchymal stem cell antigen-1 (MSCA-1) was shown to exhibit an increased osteogenic potential compared to the MSCA-1low cell fraction [1]. In order to avoid immunological reactions in future clinical studies, we established serumfree culture conditions and observed that the proliferation of the MSCA-1+ cell fraction was favored in serum-free medium [2]. We detected an earlier but weaker mineralization potential of serum-free cultured JPCs, which might significantly counter the success of future tissue engineering applications
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