Sweat Richard S, Sloas David C, Murfee Walter L
Department of Biomedical Engineering, Tulane University, New Orleans, Louisiana, USA.
Microcirculation. 2014 Aug;21(6):532-40. doi: 10.1111/micc.12132.
Lymphatic and blood microvascular systems are critical for tissue function. Insights into the coordination of both systems can be gained by investigating the relationships between lymphangiogenesis and angiogenesis. Recently, our laboratory established the rat mesentery culture model as a novel tool to investigate multicellular interactions during angiogenesis in an intact microvascular network scenario. The objective of this study was to determine whether the rat mesentery culture model can be used to study lymphangiogenesis.
Mesenteric tissue windows were harvested from adult male Wistar rats and cultured for three or five days in either serum-free MEM or MEM supplemented with VEGF-C. Tissues were immunolabeled for PECAM and LYVE-1 to identify blood and lymphatic endothelial cells, respectively. Tissues selected randomly from those containing vascular networks were quantified for angiogenesis and lymphangiogenesis.
VEGF-C treatment resulted in an increase in the density of blood vessel sprouting compared to controls by day 3. By day 5, lymphatic sprouting was increased compared to controls.
These results are consistent with in vivo findings that lymphangiogenesis lags angiogenesis after chronic stimulation and establish a tool for investigating the interrelationships between lymphangiogenesis and angiogenesis in a multisystem microvascular environment.
淋巴和血液微血管系统对组织功能至关重要。通过研究淋巴管生成与血管生成之间的关系,可以深入了解这两个系统的协调情况。最近,我们实验室建立了大鼠肠系膜培养模型,作为在完整微血管网络场景中研究血管生成过程中多细胞相互作用的新工具。本研究的目的是确定大鼠肠系膜培养模型是否可用于研究淋巴管生成。
从成年雄性Wistar大鼠获取肠系膜组织窗,在无血清MEM或补充VEGF-C的MEM中培养三或五天。分别用PECAM和LYVE-1对组织进行免疫标记,以识别血液和淋巴管内皮细胞。从含有血管网络的组织中随机选取组织,对血管生成和淋巴管生成进行定量分析。
与对照组相比,到第3天VEGF-C处理导致血管芽生密度增加。到第5天,与对照组相比,淋巴芽生增加。
这些结果与体内研究结果一致,即慢性刺激后淋巴管生成滞后于血管生成,并建立了一种在多系统微血管环境中研究淋巴管生成与血管生成之间相互关系的工具。