Brauchle Eva, Carvajal Berrio Daniel, Rieger Melanie, Schenke-Layland Katja, Reinert Siegmar, Alexander Dorothea
Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Department of Cell and Tissue Engineering, Nobelstr. 12, 70569 Stuttgart, Germany; Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany.
Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tübingen, Silcherstr. 7/1, 72076 Tübingen, Germany.
Stem Cells Int. 2017;2017:1651376. doi: 10.1155/2017/1651376. Epub 2017 Jan 23.
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.
为实现更安全的患者治疗，必须为细胞治疗建立无血清细胞培养条件。在先前的研究中，我们证明无血清培养有利于源自颌骨骨膜的MSCA-1骨祖细胞的增殖。在本研究中，使用无标记拉曼光谱法评估并比较了MSCA-1颌骨骨膜细胞（JPCs，来自3名供体）在无血清和含血清培养基条件下体外形成骨特异性基质的情况。基于标准荧光测定法，来自一名患者的JPCs在含血清培养条件下无法矿化，而其他细胞在两种条件下显示出相似的矿化水平。矿化的MSCA-1 JPCs的拉曼光谱显示，在无血清培养条件下，羟基磷灰石形成水平更高，矿物质与基质的比例更高。在含血清条件下培养的JPCs中较高的碳酸盐与磷酸盐比例和较高的结晶度表明骨形成不成熟。由于在无血清条件下胶原蛋白产生减少，与含血清条件相比，我们在胶原蛋白成熟度以及脯氨酸与羟脯氨酸比例方面获得了显著差异。我们得出结论，拉曼光谱法是评估和无创监测骨祖细胞体外矿化的有用工具。进一步的研究应扩展这一知识，并通过优化培养条件来改善JPCs的矿化。
