Embryonic rat vascular smooth muscle cells revisited - a model for neonatal, neointimal SMC or differentiated vascular stem cells?
Embryonic rat vascular smooth muscle cells revisited - a model for neonatal, neointimal SMC or differentiated vascular stem cells?
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DOI：
10.1186/2045-824x-6-6
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发表时间：
2014-03-15
期刊：
影响因子：
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通讯作者：
Cahill PA
中科院分区：
文献类型：
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作者：
Kennedy E;Hakimjavadi R;Greene C;Mooney CJ;Fitzpatrick E;Collins LE;Loscher CE;Guha S;Morrow D;Redmond EM;Cahill PA
源自胚胎大鼠胸主动脉的 A10 和 A7r5 细胞系广泛用作培养中的非分化、新生儿和新生内膜血管平滑肌细胞的模型。最近发现血管壁内存在多能血管干细胞，因此有必要重新审视这些血管细胞的身份和起源。在这种情况下，我们检查了 A10 和 A7r5 细胞系，通过确定它们的分化状态、干细胞标记物表达及其体外多能潜力，确定这些细胞系与从成年大鼠主动脉分离的多能血管干细胞之间的相似性和差异。使用免疫细胞化学、共聚焦显微镜、FACS 分析和实时定量 PCR 评估血管平滑肌细胞分化标志物（α-肌动蛋白、肌球蛋白重链、钙调蛋白）和干细胞标志物表达（Sox10、Sox17 和 S100β）。 A10和A7r5均表达血管平滑肌分化、标记物、平滑肌α-肌动蛋白、平滑肌肌球蛋白重链和钙调蛋白。在平行分析中，从大鼠主动脉外植体中分离出的多能血管干细胞的免疫细胞化学肌球蛋白重链呈阴性，但神经干细胞标记物Sox10+、神经嵴标记物、内胚层标记物Sox17+和神经胶质细胞标记物S100β+呈阳性。除了外膜祖干细胞标记、干细胞抗原-1、Sca1+ 之外，还在两种胚胎血管细胞系中检测到这种多能血管干细胞标记谱。与血清处理的细胞相比，血清剥夺导致干细胞和平滑肌细胞分化标记物表达显着增加。两种细胞类型在脂肪细胞诱导刺激后均表现出弱多能性。此外，用 DAPT 抑制 γ-分泌酶后，Notch 信号传导阻断增强了血管平滑肌和干细胞标记物的表达。我们得出的结论是，A10 和 A7r5 细胞与多能血管干细胞和外膜祖细胞具有相似的神经干细胞标记，表明新内膜干细胞来源的平滑肌细胞。这可能对其用于体外检查血管收缩和增殖表型具有重要意义。
The A10 and A7r5 cell lines derived from the thoracic aorta of embryonic rat are widely used as models of non-differentiated, neonatal and neointimal vascular smooth muscle cells in culture. The recent discovery of resident multipotent vascular stem cells within the vessel wall has necessitated the identity and origin of these vascular cells be revisited. In this context, we examined A10 and A7r5 cell lines to establish the similarities and differences between these cell lines and multipotent vascular stem cells isolated from adult rat aortas by determining their differentiation state, stem cell marker expression and their multipotency potential in vitro. Vascular smooth muscle cell differentiation markers (alpha-actin, myosin heavy chain, calponin) and stem cell marker expression (Sox10, Sox17 and S100β) were assessed using immunocytochemistry, confocal microscopy, FACS analysis and real-time quantitative PCR. Both A10 and A7r5 expressed vascular smooth muscle differentiation, markers, smooth muscle alpha - actin, smooth muscle myosin heavy chain and calponin. In parallel analysis, multipotent vascular stem cells isolated from rat aortic explants were immunocytochemically myosin heavy chain negative but positive for the neural stem cell markers Sox10+, a neural crest marker, Sox17+ the endoderm marker, and the glia marker, S100β+. This multipotent vascular stem cell marker profile was detected in both embryonic vascular cell lines in addition to the adventitial progenitor stem cell marker, stem cell antigen-1, Sca1+. Serum deprivation resulted in a significant increase in stem cell and smooth muscle cell differentiation marker expression, when compared to serum treated cells. Both cell types exhibited weak multipotency following adipocyte inductive stimulation. Moreover, Notch signaling blockade following γ-secretase inhibition with DAPT enhanced the expression of both vascular smooth muscle and stem cell markers. We conclude that A10 and A7r5 cells share similar neural stem cell markers to both multipotent vascular stem cells and adventitial progenitors that are indicative of neointimal stem-derived smooth muscle cells. This may have important implications for their use in examining vascular contractile and proliferative phenotypes in vitro.