Perfused 3D angiogenic sprouting in a high-throughput in vitro platform.
Perfused 3D angiogenic sprouting in a high-throughput in vitro platform.
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在高通量的体外平台中灌注3D血管生成。
DOI：
10.1007/s10456-018-9647-0
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发表时间：
2019-03
期刊：
影响因子：
9.8
通讯作者：
Hankemeier T
中科院分区：
文献类型：
--
作者：
van Duinen V;Zhu D;Ramakers C;van Zonneveld AJ;Vulto P;Hankemeier T
血管生成萌芽，即新血管从预先存在的血管生长，是由细胞微环境内的线索（如生化梯度和灌注）协调的。然而，这些线索中的许多在目前的血管生成发芽的体外模型中缺失。我们在这里描述了一个体外平台，它集成了灌注和稳定的生物分子梯度的产生，并证明了它的潜力，以研究更多的生理相关的血管生成发芽和微血管稳定。该平台由40个可单独寻址的微流体单元阵列组成，这些单元能够针对三维胶原蛋白-1基质培养灌注的微血管。在引入促血管生成因子的梯度后，内皮细胞分化成侵入基质的尖端细胞。持续暴露导致柄细胞持续迁移并形成管腔。血管内皮生长因子-165（VEGF-165）、佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）和鞘氨醇-1-磷酸酯（S1 P）的组合是触发稳健、定向血管生成的最佳鸡尾酒，其中S1 P对于指导和重复芽形成至关重要。长时间的暴露迫使血管新生芽通过胶原蛋白到达另一个通道。这导致胶原内血管生成芽的重塑：与其他灌注通道结合的血管生成芽保持稳定，而那些没有收缩和降解的血管生成芽。此外，用150 kDa FITC-葡聚糖灌注显示，虽然血管生成芽最初是渗漏的，但一旦它们完全穿过胶原泳道，它们就变得不渗漏。这表明，一旦发生吻合，芽成熟，并表明灌注可以作为一个重要的存活和稳定因素的血管生成微血管。该平台的稳健性与包括更生理相关的三维微环境的可能性相结合，使我们的平台特别适合于研究体外血管生成。本文的在线版本（10.1007/s10456-018-9647-0）包含补充材料，可供授权用户使用。
Angiogenic sprouting, the growth of new blood vessels from pre-existing vessels, is orchestrated by cues from within the cellular microenvironment, such as biochemical gradients and perfusion. However, many of these cues are missing in current in vitro models of angiogenic sprouting. We here describe an in vitro platform that integrates both perfusion and the generation of stable biomolecular gradients and demonstrate its potential to study more physiologically relevant angiogenic sprouting and microvascular stabilization. The platform consists of an array of 40 individually addressable microfluidic units that enable the culture of perfused microvessels against a three-dimensional collagen-1 matrix. Upon the introduction of a gradient of pro-angiogenic factors, the endothelial cells differentiated into tip cells that invaded the matrix. Continuous exposure resulted in continuous migration and the formation of lumen by stalk cells. A combination of vascular endothelial growth factor-165 (VEGF-165), phorbol 12-myristate 13-acetate (PMA), and sphingosine-1-phosphate (S1P) was the most optimal cocktail to trigger robust, directional angiogenesis with S1P being crucial for guidance and repetitive sprout formation. Prolonged exposure forces the angiogenic sprouts to anastomose through the collagen to the other channel. This resulted in remodeling of the angiogenic sprouts within the collagen: angiogenic sprouts that anastomosed with the other perfusion channel remained stable, while those who did not retracted and degraded. Furthermore, perfusion with 150 kDa FITC-Dextran revealed that while the angiogenic sprouts were initially leaky, once they fully crossed the collagen lane they became leak tight. This demonstrates that once anastomosis occurred, the sprouts matured and suggests that perfusion can act as an important survival and stabilization factor for the angiogenic microvessels. The robustness of this platform in combination with the possibility to include a more physiological relevant three-dimensional microenvironment makes our platform uniquely suited to study angiogenesis in vitro. The online version of this article (10.1007/s10456-018-9647-0) contains supplementary material, which is available to authorized users.
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影响因子：
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作者：
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作者：
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通讯作者：
Hla, Timothy
DOI：
10.1007/978-1-62703-604-7_2
发表时间：
2013-01-01
期刊：
CELL-CELL INTERACTIONS: METHODS AND PROTOCOLS, 2ND EDITION
影响因子：
--
作者：
Davis, George E.;Kim, Dae Joong;Stratman, Amber N.
通讯作者：
Stratman, Amber N.