An angiogenesis model for investigating multicellular interactions across intact microvascular networks
An angiogenesis model for investigating multicellular interactions across intact microvascular networks
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DOI：
10.1152/ajpheart.00552.2012
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发表时间：
2013-01-01
影响因子：
4.8
通讯作者：
Murfee, Walter L.
中科院分区：
文献类型：
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作者：
Stapor, Peter C.;Azimi, Mohammad S.;Murfee, Walter L.
Stapor PC，Azimi MS，Ahsan T，Murfee WL.一个研究完整微血管网络中多细胞相互作用的血管生成模型。Am J Physiol Heart Circ Physiol 304：H235-H245，2013。首次发表于2012年11月2日; doi：10.1152/ajpheart.00552.2012.-开发旨在操纵微血管重塑的治疗方法需要更好地理解血管生成以及血管生成如何与其他网络重塑过程（如淋巴管生成和神经发生）相关。本研究的目的是开发一种血管生成模型，能够在分子水平上探测完整的成人微血管网络中的多细胞和多系统相互作用。无菌收获成年雄性Wistar大鼠肠系膜窗，并在无血清最低必需培养基中培养。活力/细胞毒性分析显示细胞保持存活至少7天。免疫组织化学标记血小板内皮细胞粘附分子（PECAM），神经元-胶质细胞抗原2（NG 2），淋巴管内皮透明质酸受体-1（LYVE-1），和III类β-微管蛋白，分别确定内皮细胞，周细胞，神经，和神经。补充有bFGF或VEGF的培养基诱导内皮细胞从现有血管出芽的增加。NG 2功能性阻断抗体靶向周细胞抑制了两个生长因子组的内皮细胞发芽。与单独培养基或bFGF组相比，VEGF引起淋巴/血液内皮细胞连接数量增加。最后，在通过补充具有20%血清的培养基刺激血管生成之前和之后的相同网络的比较鉴定了断开的内皮段重新连接到附近血管的能力。结果建立了一种新的原位血管生成模型，用于研究完整网络内毛细血管发芽的位置，周细胞的作用，淋巴/血液内皮细胞的相互作用，以及特定内皮细胞片段的命运。大鼠肠系膜培养系统提供了一个独特的工具，了解复杂的动态与血管生成在一个完整的成人组织。
Stapor PC, Azimi MS, Ahsan T, Murfee WL. An angiogenesis model for investigating multicellular interactions across intact microvascular networks. Am J Physiol Heart Circ Physiol 304: H235-H245, 2013. First published November 2, 2012; doi:10.1152/ajpheart.00552.2012.-Developing therapies aimed at manipulating microvascular remodeling requires a better understanding of angiogenesis and how angiogenesis relates to other network remodeling processes, such as lymphangiogenesis and neurogenesis. The objective of this study was to develop an angiogenesis model that enables probing of multicellular and multisystem interactions at the molecular level across an intact adult microvascular network. Adult male Wistar rat mesenteric windows were aseptically harvested and cultured in serum-free minimum essential media. Viability/cytotoxicity analysis revealed that cells remain alive for at least 7 days. Immunohistochemical labeling at 3 days for platelet endothelial cell adhesion molecule (PECAM), neuron-glial antigen 2 (NG2), lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), and class III beta-tubulin identified endothelial cells, pericytes, lymphatics, and nerves, respectively. Media supplemented with bFGF or VEGF induced an increase in endothelial cell sprouting off existing vessels. Endothelial cell sprouting in both growth factor groups was inhibited by targeting pericytes with NG2 functional blocking antibody. VEGF caused an increase in the number of lymphatic/blood endothelial cell connections compared with media alone or bFGF groups. Finally, the comparison of the same network before and after angiogenesis stimulated by the supplement of media with 20% serum identified the ability of disconnected endothelial segments to reconnect to nearby vessels. The results establish a novel in situ angiogenesis model for investigating the location of capillary sprouting within an intact network, the role of pericytes, lymphatic/blood endothelial cell interactions, and the fate of specific endothelial cell segments. The rat mesentery culture system offers a unique tool for understanding the complex dynamics associated with angiogenesis in an intact adult tissue.