Human and Swine Dental Pulp Stem Cells Form a Vascularlike Network after Angiogenic Differentiation in Comparison with Endothelial Cells: A Quantitative Analysis.
Human and Swine Dental Pulp Stem Cells Form a Vascularlike Network after Angiogenic Differentiation in Comparison with Endothelial Cells: A Quantitative Analysis.
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DOI：
10.1016/j.joen.2016.11.015
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发表时间：
2017-04
影响因子：
4.2
通讯作者：
Huang GT
中科院分区：
文献类型：
--
作者：
Aksel H;Huang GT
目的是量化人和猪牙髓干细胞（DPSC）与内皮细胞相比在血管生成诱导后的血管网络形成能力。在内皮生长培养基（EGM）中诱导原代人（h）DPSC或猪（s）DPSC 7天。免疫组化法检测血管内皮标志物血管性血友病因子（vWF）的表达。将诱导的（iDPSC）和未诱导的（n-iDPSC）以不同的细胞数接种到Matrigel上用于血管网络形成测定，并在4、8、12和18小时后与人内皮细胞（hMEC）相比进行分析。使用ImageJ进行血管小管形成的定量分析。还通过n-iDPSC和iDPSC的共培养进行血管网络形成。通过免疫荧光检测n-iDPSC和iDPSC（人和猪）中的vWF。延时显微镜观察显示血管网由iDPSCs形成，而非n-iDPSCs。4 h后，iDPSCs显示血管网络形成，而FDPSC开始聚集并形成簇。基于定量分析，与hMEC相比，ihDPSC显示出在基质胶中形成血管网络的类似能力。与ihDPSC或hMEC相比，isDPSC在形成包括段、节点和网格的网络结构方面具有更高的能力（p<0.05）。isDPSC比ihDPSC和hMEC更显著（p<0.05）。共培养实验显示n-ihDPSCs共定位于由ihDPSCs形成的血管生成小管和血管网络上。我们的研究结果表明，iDPSCs与其非诱导对应物的组合可用作未来的临床策略，用于在牙髓-牙本质再生过程中增强血管生成。
The aim was to quantify vascular network formation capacity after angiogenic induction of human and swine dental pulp stem cells (DPSCs) in comparison to endothelial cells. Primary human (h) DPSCs or swine (s) DPSCs were induced in endothelial growth medium (EGM) for 7 days. The expression of endothelial marker, von Willebrand Factor (vWF) was determined by immunostaining. Induced (iDPSCs) and non-induced (n-iDPSCs) were seeded at different cell numbers onto Matrigel for vascular network formation assays and analyzed after 4, 8, 12, and 18 h in comparison to human endothelial cells (hMECs). Quantitative analysis of vascular tubule formation was performed using ImageJ. The vascular network formation was also conducted by co-culturing of n-iDPSCs and iDPSCs. vWF was detected by immunofluorescence in both n-iDPSCs and iDPSCs (human and swine). Time-lapse microscopic observation showed that the vascular network was formed by iDPSCs, but not n-iDPSCs. After 4 h, iDPSCs showed vascular network formation while FDPSCs started to aggregate and formed clusters. ihDPSCs displayed a similar capacity to form vascular networks in Matrigel compared to hMECs based on quantitative analysis. isDPSCs had a higher capacity compared to ihDPSCs or hMECs (p<0.05) in forming the network structures including segments, nodes and mesh. isDPSCs than ihDPSCs and hMECs (p<0.05). Co-culture experiment showed that n-ihDPSCs co-localized on the angiogenic tubules and vascular networks formed by ihDPSCs. Our findings indicate that iDPSCs in combination of their non-induced counterparts may be utilized as a future clinical strategy for enhancing angiogenesis during the process of pulp-dentin regeneration.