Comparison of three in vitro human 'angiogenesis' assays with capillaries formed in vivo
Comparison of three in vitro human 'angiogenesis' assays with capillaries formed in vivo
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DOI：
10.1023/a:1012218401036
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发表时间：
2001-01-01
期刊：
影响因子：
9.8
通讯作者：
Lewis, C. E.
中科院分区：
文献类型：
--
作者：
Donovan, D.;Brown, N. J.;Lewis, C. E.
血管生成检测是研究血管生成机制和调节新生血管形成的治疗策略的潜在发展的重要工具。体内血管生成分析被认为是其中最有信息量的，但通常昂贵、耗时，而且需要专家培训才能进行。体外分析往往更快速、更便宜、更容易解释。在体外，血管生成试验的原理是内皮细胞在支持性基质上培养时形成管状结构。涉及小鼠肿瘤基质Matrigel(或生长因子降低形式)的分析现在是最常见的体外小管形成分析方法。然而，最近发展了另一种小管形成试验，其中内皮细胞与成纤维细胞共培养。在这里，我们使用定量图像分析来比较Matrigel实验和这种新的‘共培养’实验中形成的小管的形态特征，以及体内微血管床中形成的毛细血管的形态特征。在标准的和生长因子减少的Matrigel实验中形成的小管较短且相对均匀，而在共培养实验中形成的小管明显更不均匀，由短和长相互连接的小管组成，这些小管比Matrigel小管更接近毛细血管。此外，Matrigel上的细胞，以及较小程度的生长因子减少(GFR)Matrigel上的细胞，经常聚集成大的细胞聚集体，这在共培养试验中很少见。此外，我们证明Matrigel通过各种非内皮细胞类型刺激小管形成，这表明内皮细胞的小管形成可能并不代表这种细胞类型的真正分化。总之，共培养试验中的小管形态似乎更能代表体内毛细血管的形成，而不是Matrigel试验中任一种形式的内皮细胞的变化。
Angiogenesis assays are an important tool for studying both the mechanisms of angiogenesis and the potential development of therapeutic strategies to modulate neovascularisation. In vivo angiogenesis assays are considered to be the most informative of these but are often expensive, time-consuming and require specialist training to perform. In vitro assays tend to be more rapid, less expensive and easier to interpret. In vitro angiogenesis assays operate on the principle that endothelial cells form tubule-like structures when cultured on a supportive matrix. Assays involving a matrix derived from murine tumours, Matrigel (or a growth factor reduced form of this), are now the most common in vitro tubule formation assays. However, another tubule formation assay has recently been developed in which endothelial cells are co-cultured with fibroblasts. Here, we have used quantitative image analysis to compare the morphological features of tubules formed in the Matrigel assay and this new 'Co-culture' assay, with those of capillaries formed in a microvascular bed in vivo. Tubules formed in standard and growth factor reduced Matrigel assays were short and relatively homogeneous, whereas those formed in the Co-culture assay were significantly more heterogeneous, consisting of both short and long interconnecting tubules that more closely resembled capillaries than Matrigel tubules. Moreover, cells on Matrigel, and to a lesser extent growth factor reduced (GFR) Matrigel, often clumped into large cell aggregates, a feature rarely seen in the Co-culture assay. In addition, we demonstrate that Matrigel stimulates tubule formation by various non-endothelial cell types, suggesting that tubule formation by endothelial cells may not represent true differentiation of this cell type. In summary, the morphology of tubules in the Co-culture assay appears more representative of capillary formation in vivo, than the endothelial cell changes that occur in either form of Matrigel assay.