Background

Neurovascular patterning is an emerging area of microvascular research. While overlapping molecular signals highlight links between angiogenesis and neurogenesis, advancing our understanding is limited by a lack of in vitro models containing both systems. One potential model is the rat mesentery culture model, which our laboratory has recently introduced as an ex vivo tool to investigate cellular dynamics during angiogenesis in a microvascular network scenario. The objective of this study was to demonstrate the use of the rat mesentery culture model as an ex vivo platform for maintaining the spatiotemporal relationship between blood vessels and peripheral nerves during angiogenesis in adult microvascular networks.

Methods

Adult male Wistar rat mesenteric tissue windows were harvested, rinsed in sterile DPBS and medium and then cultured per group: 1) MEM alone and 2) NBM with NGF and 20 % FBS (nerve culture medium). On day 3 post culture tissues were labeled for endothelial (PECAM) and neural (class III β-tubulin, NG2, tyrosine hydroxylase, CGRP) markers.

Results

In MEM alone tissues nerve segment degeneration was supported by discontinuous nerve or absence of nerve marker labeling. Nerve presence at the arteriole level and capillary level was maintained for the nerve culture medium group compared to day 0, non-cultured control group (unstimulated).

Comparison with existing methods and conclusions

The results support the use of specific medium types to maintain nerve presence across cultured microvascular networks and implicates the rat mesentery culture model as a novel ex vivo tool for investigating neurovascular patterning in adult tissues.

中文翻译：

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大鼠肠系膜培养模型中成人周围神经和微血管网络的维护。

背景

神经血管模式是微血管研究的一个新兴领域。虽然重叠的分子信号突出了血管生成和神经发生之间的联系，但由于缺乏包含这两种系统的体外模型，我们的理解受到了限制。一个潜在的模型是大鼠肠系膜培养模型，我们的实验室最近将其作为一种体外工具引入，用于研究微血管网络场景中血管生成过程中的细胞动力学。本研究的目的是证明使用大鼠肠系膜培养模型作为体外平台，在成人微血管网络的血管生成过程中维持血管和周围神经之间的时空关系。

方法

收获成年雄性 Wistar 大鼠肠系膜组织窗，在无菌 DPBS 和培养基中冲洗，然后每组培养：1) 单独的 MEM 和 2) 具有 NGF 和 20% FBS（神经培养基）的 NBM。在培养后第 3 天，对组织进行内皮 (PECAM) 和神经（III 类 β-微管蛋白、NG2、酪氨酸羟化酶、CGRP）标记。

结果

在单独的 MEM 组织中，神经节段退化由不连续的神经或没有神经标记标记支持。与第 0 天的非培养对照组（未刺激）相比，神经培养基组在小动脉水平和毛细血管水平上保持神经存在。

与现有方法和结论的比较

结果支持使用特定的培养基类型来维持培养的微血管网络中的神经存在，并将大鼠肠系膜培养模型作为一种新的体外工具，用于研究成人组织中的神经血管模式。