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Abstract：
Vascular smooth muscle cells (SMCs) from distinct anatomic locations derive from different embryonic origins. Here we investigated the respective potential of different embryonic origin-specific SMCs derived from human embryonic stem cells (hESCs) to support endothelial network formation in vitro. SMCs of three distinct embryological origins were derived from an mStrawberry-expressing hESC line and were cocultured with green fluorescent protein-expressing human umbilical vein endothelial cells (HUVECs) to investigate the effects of distinct SMC subtypes on endothelial network formation. Quantitative analysis demonstrated that lateral mesoderm (LM)-derived SMCs best supported HUVEC network complexity and survival in three-dimensional coculture in Matrigel. The effects of the LM-derived SMCs on HUVECs were at least in part paracrine in nature. A TaqMan array was performed to identify the possible mediators responsible for the differential effects of the SMC lineages, and a microarray was used to determine lineage-specific angiogenesis gene signatures. Midkine (MDK) was identified as one important mediator for the enhanced vasculogenic potency of LM-derived SMCs. The functional effects of MDK on endothelial network formation were then determined by small interfering RNA-mediated knockdown in SMCs, which resulted in impaired network complexity and survival of LM-derived SMC cocultures. The present study is the first to show that SMCs from distinct embryonic origins differ in their ability to support HUVEC network formation. LM-derived SMCs best supported endothelial cell network complexity and survival in vitro, in part through increased expression of MDK. A lineage-specific approach might be beneficial for vascular tissue engineering and therapeutic revascularization.
CONCLUSIONS: Mural cells are essential for the stabilization and maturation of new endothelial cell networks. However, relatively little is known of the effect of the developmental origins of mural cells on their signaling to endothelial cells and how this affects vessel development. The present study demonstrated that human smooth muscle cells (SMCs) from distinct embryonic origins differ in their ability to support endothelial network formation. Lateral mesoderm-derived SMCs best support endothelial cell network complexity and survival in vitro, in part through increased expression of midkine. A lineage-specific approach might be beneficial for vascular tissue engineering and therapeutic revascularization.
CONCLUSIONS: Mural cells are essential for the stabilization and maturation of new endothelial cell networks. However, relatively little is known of the effect of the developmental origins of mural cells on their signaling to endothelial cells and how this affects vessel development. The present study demonstrated that human smooth muscle cells (SMCs) from distinct embryonic origins differ in their ability to support endothelial network formation. Lateral mesoderm-derived SMCs best support endothelial cell network complexity and survival in vitro, in part through increased expression of midkine. A lineage-specific approach might be beneficial for vascular tissue engineering and therapeutic revascularization.
摘要：
来自不同解剖位置的血管平滑肌细胞（SMC）来自不同的胚胎起源。在这里，我们研究了来自人胚胎干细胞（hESC）的不同胚胎起源特异性SMC各自的潜力，以支持体外内皮细胞网络形成。三种不同胚胎学起源的SMC来自表达mStrawberry的hESC系，并与表达绿色荧光蛋白的人脐静脉内皮细胞（HUVEC）共培养，以研究不同SMC亚型对内皮网络形成的影响。定量分析表明，外侧中胚层（LM）衍生的SMC在Matrigel的三维共培养中最能支持HUVEC网络的复杂性和存活。LM衍生的SMC对HUVEC的作用在本质上至少部分是旁分泌的。进行TaqMan阵列以鉴定负责SMC谱系差异效应的可能介体,和微阵列用于确定谱系特异性血管生成基因特征。Midkine（MDK）被认为是LM衍生的SMC增强血管生成能力的重要介质。然后通过小干扰RNA介导的SMC敲低来确定MDK对内皮细胞网络形成的功能影响，这导致LM衍生的SMC共培养物的网络复杂性和存活受损。本研究首次表明来自不同胚胎起源的SMC在支持HUVEC网络形成的能力上有所不同。LM衍生的SMC在体外最支持内皮细胞网络复杂性和存活，部分通过增加MDK的表达。谱系特异性方法可能对血管组织工程和治疗性血运重建有益。
结论：壁细胞对于新的内皮细胞网络的稳定和成熟至关重要。然而,对于壁细胞的发育起源对其向内皮细胞的信号传导的影响以及这如何影响血管发育，人们知之甚少。本研究表明，来自不同胚胎起源的人平滑肌细胞（SMC）在支持内皮细胞网络形成的能力上有所不同。外侧中胚层来源的SMC最好地支持内皮细胞网络复杂性和体外存活，部分通过增加中期因子的表达。谱系特异性方法可能对血管组织工程和治疗性血运重建有益。
结论：壁细胞对于新的内皮细胞网络的稳定和成熟至关重要。然而,对于壁细胞的发育起源对其向内皮细胞的信号传导的影响以及这如何影响血管发育，人们知之甚少。本研究表明，来自不同胚胎起源的人平滑肌细胞（SMC）在支持内皮细胞网络形成的能力上有所不同。外侧中胚层来源的SMC最好地支持内皮细胞网络复杂性和体外存活，部分通过增加中期因子的表达。谱系特异性方法可能对血管组织工程和治疗性血运重建有益。
