1 Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy.
2 Laboratory of Experimental Medicine and Environmental Pathology, Rieti University Hub "Sabina Universitas," Rieti, Italy.
Stem Cells Dev. 2019 May 15;28(10):695-706. doi: 10.1089/scd.2018.0192. Epub 2019 Apr 23.
Dental pulp has been revealed as an accessible and a rich source of mesenchymal stem cells (MSCs) and its biological potential is currently under intense investigation. MSCs from dental pulp stem cells (DPSCs) have been indicated as a heterogeneous population oriented not only in repairing dentine but also in maintaining vascular and nervous homeostasis of the teeth. We sought to verify the phenotype of cells isolated from dental pulp of young donors and to investigate in vitro their role as pericyte-like cells. Specifically, we evaluated how culture conditions can modulate expression of pericyte markers in DPSCs and their capacity to stabilize endothelial tubes in vitro. DPSCs cultured in standard conditions expressed MSC markers and demonstrated to contain a population expressing the pericyte marker NG2. These DPSCs were associated with low sprouting capacity in extra-cellular (EC) Matrix and limited ability in retaining tubes formed by endothelial cells in a coculture angiogenesis model. When cultured in endothelial growth medium (EGM)-2, DPSCs significantly upregulated NG2, and partially alpha-smooth muscle actin. The resulting population conserved the stem marker CD73, but was negative for calponin and endothelial markers. EGM-2-conditioned DPSCs showed a higher sprouting ability in EC Matrix and efficient association with human umbilical vein endothelial cells allowing the partial retention of endothelial tubes for several days. Among growth factors contained in EGM-2 we identified basic fibroblast growth factor (bFGF) as mainly responsible for NG2 upregulation and long-term stabilization of endothelial tubes. According to the in vitro analysis, DPSCs represent an effective source of pericytes and the appropriate culture conditions could result in a population with a promising ability to stabilize vessels and promote vascular maturation.
牙髓已被证明是间充质干细胞(MSCs)的一个可及且丰富的来源,其生物学潜能目前正受到广泛研究。牙髓干细胞(DPSCs)来源的 MSCs 已被证明是一种异质性群体,不仅在修复牙本质方面具有潜力,而且在维持牙齿的血管和神经内稳态方面也具有潜力。我们试图验证从小龄供体的牙髓中分离出的细胞的表型,并研究其在体外作为周细胞样细胞的作用。具体而言,我们评估了培养条件如何调节 DPSCs 中周细胞标志物的表达及其在体外稳定内皮管的能力。在标准条件下培养的 DPSCs 表达 MSC 标志物,并证明含有表达周细胞标志物 NG2 的群体。这些 DPSCs 与细胞外基质(EC 基质)中低发芽能力和在共培养血管生成模型中保留内皮细胞形成的管的能力有限相关。当在内皮细胞生长培养基(EGM-2)中培养时,DPSCs 显著上调 NG2,并部分上调 alpha-平滑肌肌动蛋白。由此产生的群体保留了干细胞标志物 CD73,但阴性表达钙调蛋白和内皮标志物。在 EGM-2 条件下培养的 DPSCs 在 EC 基质中具有更高的发芽能力,并与脐静脉内皮细胞有效关联,允许内皮管在数天内部分保留。在 EGM-2 中包含的生长因子中,我们确定碱性成纤维细胞生长因子(bFGF)主要负责 NG2 的上调和内皮管的长期稳定。根据体外分析,DPSCs 是周细胞的有效来源,适当的培养条件可能会产生一种具有稳定血管和促进血管成熟的有前途能力的群体。
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