Notch Signaling Pathway Regulates Angiogenesis via Endothelial Cell in 3D Co-Culture Model.
Notch Signaling Pathway Regulates Angiogenesis via Endothelial Cell in 3D Co-Culture Model.
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Notch 信号通路在 3D 共培养模型中通过内皮细胞调节血管生成
DOI：
10.1002/jcp.25681
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
林云锋
中科院分区：
文献类型：
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作者：
Zhao Dan;Xue Changyue;Lin Shiyu;Shi Sirong;Li Qianshun;Liu Mengting;Cai Xiaoxiao;林云锋
本研究旨在研究Notch信号通路在脂肪源性基质细胞（ASC）和内皮细胞（EC）共培养的三维（3D）胶原凝胶模型中对血管生成的作用。将绿色荧光蛋白标记的小鼠ASCs和红色荧光蛋白标记的ECs分别植入体外构建三维胶原凝胶模型，采用激光共聚焦显微镜观察Notch信号诱导剂Jagged 1、抑制剂DAPT和PBS对ASCs和ECs血管生成的影响。半定量PCR和免疫荧光染色检测血管生成相关基因和蛋白的表达。与对照组相比，Jagged 1处理促进了共培养凝胶中的血管生成，而DAPT处理减弱了血管生成。在ASCs和ECs共培养体系中，Jagged 1处理可增加ECs中VEGFA、VEGFB、Notch 1、Notch 2、Hes 1、Hey 1、VEGFR 1的基因表达，而DAPT处理可降低ECs中VEGFA、VEGFB、Notch 1、Hes 1、Hey 1的蛋白表达。而VEGFR 3则相反。然而，同样的结果并没有完全出现在ASCs。这些结果表明，当ASC和EC在3D胶原凝胶模型中共培养时，VEGFA/B-Notch 1/2-Hes 1/Hey 1-VEGFR 1/3信号轴在血管生成中起重要作用。J.细胞。232：1548-1558，2017。© 2016 Wiley Periodicals，Inc.
This study aimed to investigate the role of Notch signaling pathway for angiogenesis in a three‐dimensional (3D) collagen gel model with co‐culture of adipose‐derived stromal cells (ASCs) and endothelial cells (ECs). A 3D collagen gel model was established in vitro by implanting both ASCs from green fluorescent protein‐labeled mouse and ECs from red fluorescent protein‐labeled mouse, and the phenomena of angiogenesis with Notch signaling inducer Jagged1, inhibitor DAPT and PBS, respectively were observed by confocal laser scanning microscopy. Semi‐quantitative PCR and immunofluorescent staining were conducted to detect expressions of angiogenesis‐related genes and proteins. Angiogenesis in the co‐culture gels was promoted by Jagged1 treatment while attenuated by DAPT treatment, compared to control group. In co‐culture system of ASCs and ECs, the gene expressions ofVEGFA, VEGFB, Notch1, Notch2, Hes1, Hey1, VEGFR1,and the protein expression of VEGFA, VEGFB, Notch1, Hes1, Hey1 were increased by Jagged1 treatment and decreased by DAPT treatment in ECs. And the result ofVEGFR3was the opposite. However, the same results did not appear completely in ASCs. These results revealed the VEGFA/B‐Notch1/2‐Hes1/Hey1‐ VEGFR1/3 signal axis played an important role in angiogenesis when ASCs and ECs were co‐cultured in a 3D collagen gel model. J. Cell. Physiol. 232: 1548–1558, 2017. © 2016 Wiley Periodicals, Inc.