The effect of retinal pigment epithelial cell patch size on growth factor expression
The effect of retinal pigment epithelial cell patch size on growth factor expression
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DOI：
10.1016/j.biomaterials.2014.01.016
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发表时间：
2014-04-01
期刊：
影响因子：
14
通讯作者：
Collier, Charles Patrick
中科院分区：
文献类型：
--
作者：
Vargis, Elizabeth;Peterson, Cristen B.;Collier, Charles Patrick
使用微图案化技术控制培养中生长的视网膜色素上皮（RPE）细胞的空间组织，以检查斑块大小对细胞健康和分化的影响。理解这种效应是开发多路复用高通量流体测定的关键步骤，并提供了用于复制与年龄相关性黄斑变性（AMD）期间视网膜组织恶化相关的疾病状态的模型。纤连蛋白在聚苯乙烯和玻璃基底上的微接触印刷用于促进细胞附着，形成受控尺寸和形状的RPE贴片。这些集落模拟在退行性疾病如AMD期间视网膜中发生的萎缩和功能丧失的效果。在细胞生长72小时后，测量血管内皮生长因子（VEGF）（AMD的重要生物标志物）的水平。对细胞进行计数，并通过荧光显微镜评估细胞活力和紧密连接形成的形态学指标。随着集落大小的减小，测量到每个细胞的VEGF表达增加高达两倍，这表明RPE细胞的局部微环境和细胞间的连接影响生长因子表达，导致疾病如AMD的开始和进展。(c)2014爱思唯尔有限公司版权所有。
The spatial organization of retinal pigment epithelial (RPE) cells grown in culture was controlled using micropatterning techniques in order to examine the effect of patch size on cell health and differentiation. Understanding this effect is a critical step in the development of multiplexed high throughput fluidic assays and provides a model for replicating disease states associated with the deterioration of retinal tissue during age-related macular degeneration (AMD). Microcontact printing of fibronectin on polystyrene and glass substrates was used to promote cell attachment, forming RPE patches of controlled size and shape. These colonies mimic the effect of atrophy and loss-of-function that occurs in the retina during degenerative diseases such as AMD. After 72 h of cell growth, levels of vascular endothelial growth factor (VEGF), an important biomarker of AMD, were measured. Cells were counted and morphological indicators of cell viability and tight junction formation were assessed via fluorescence microscopy. Up to a twofold increase of VEGF expression per cell was measured as colony size decreased, suggesting that the local microenvironment of, and connections between, RPE cells influences growth factor expression leading to the initiation and progression of diseases such as AMD. (c) 2014 Elsevier Ltd. All rights reserved.