Osteogenic Differentiation of Mesenchymal Stem Cells on Pregenerated Extracellular Matrix Scaffolds in the Absence of Osteogenic Cell Culture Supplements
Osteogenic Differentiation of Mesenchymal Stem Cells on Pregenerated Extracellular Matrix Scaffolds in the Absence of Osteogenic Cell Culture Supplements
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DOI：
10.1089/ten.tea.2009.0583
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发表时间：
2010-02-01
影响因子：
4.1
通讯作者：
Kasper, F. Kurtis
中科院分区：
文献类型：
--
作者：
Thibault, Richard A.;Baggett, L. Scott;Kasper, F. Kurtis
本研究采用全因素方差设计，研究了四个因素：全骨髓细胞、体外矿化细胞外基质(ECM)的存在、地塞米松的存在以及培养时间的变化对在聚合物支架上培养的间充质干细胞(MSCs)增殖和成骨分化的影响。将电纺聚己内酯(PCL)纤维网状支架与大鼠骨髓间充质干细胞复合，在完全成骨介质中培养12天，形成含有矿化细胞外基质的支架。将MSCs或MSCs和整个骨髓细胞接种到去细胞的ECM支架(PCL/ECM)或普通的PCL支架上，在含有或不含地塞米松的培养液中静态培养4、8和16天。在每个培养周期后，通过DNA分析确定细胞数量，通过碱性磷酸酶活性和钙离子测定确定细胞的成骨分化状态。将MSCs接种于PCL/ECM构建物上，在含或不含地塞米松的培养液中培养16天后，观察到相同数量的钙沉积。当整个骨髓细胞与MSCs在不含地塞米松的PCL支架上共培养时，随着时间的推移，细胞数量明显增加。这项研究证实，即使在没有地塞米松的情况下，种植到含有ECM的构建物上的MSCs仍能保持成骨分化，并且MSCs与没有地塞米松和ECM的全骨髓细胞共培养可以促进整个骨髓中存在的一个或多个细胞群的增殖。
This study utilized a full-factorial design to investigate the effect of four factors: presence of whole bone marrow cells, presence of in vitro-generated mineralized extracellular matrix (ECM), presence of dexamethasone, and variations in culture duration, on the proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) cultured on a polymer scaffold. Electrospun poly(e-caprolactone) (PCL) fiber mesh scaffolds were seeded with rat MSCs and cultured in complete osteogenic medium for 12 days to generate constructs containing mineralized ECM. MSCs or MSCs and whole bone marrow cells were seeded onto decellularized ECM constructs (PCL/ECM) or plain PCL scaffolds and cultured statically for 4, 8, and 16 days in medium either with or without dexamethasone. After each culture period, the cell number was determined by DNA analysis, and the osteogenic differentiation state of the cells was determined by alkaline phosphatase activity and calcium assays. MSCs seeded onto PCL/ECM constructs and cultured in medium either with or without dexamethasone demonstrated similar amounts of calcium deposition after 16 days. A significant increase in cell number over time compared with all other groups was observed when whole bone marrow cells were cocultured with MSCs on PCL scaffolds in medium without dexamethasone. This study establishes that the osteogenic differentiation of MSCs seeded onto ECM-containing constructs is maintained even in the absence of dexamethasone and that the coculture of MSCs and whole bone marrow cells without dexamethasone and ECM enhances the proliferation of a cell population (or populations) present in the whole bone marrow.