Blackstone Britani N, Palmer Andre F, Rilo Horacio R, Powell Heather M
1 Department of Biomedical Engineering, The Ohio State University , Columbus, Ohio.
Tissue Eng Part A. 2014 Jul;20(13-14):1784-93. doi: 10.1089/ten.TEA.2013.0107. Epub 2014 Feb 24.
Recently, in vitro diagnostic tools have shifted focus toward personalized medicine by incorporating patient cells into traditional test beds. These cell-based platforms commonly utilize two-dimensional substrates that lack the ability to support three-dimensional cell structures seen in vivo. As monolayer cell cultures have previously been shown to function differently than cells in vivo, the results of such in vitro tests may not accurately reflect cell response in vivo. It is therefore of interest to determine the relationships between substrate architecture, cell structure, and cell function in 3D cell-based platforms. To investigate the effect of substrate architecture on insulinoma organization and function, insulinomas were seeded onto 2D gelatin substrates and 3D fibrous gelatin scaffolds with three distinct fiber diameters and fiber densities. Cell viability and clustering was assessed at culture days 3, 5, and 7 with baseline insulin secretion and glucose-stimulated insulin production measured at day 7. Small, closely spaced gelatin fibers promoted the formation of large, rounded insulinoma clusters, whereas monolayer organization and large fibers prevented cell clustering and reduced glucose-stimulated insulin production. Taken together, these data show that scaffold properties can be used to control the organization and function of insulin-producing cells and may be useful as a 3D test bed for diabetes drug development.
最近,体外诊断工具通过将患者细胞纳入传统测试平台,已将重点转向个性化医疗。这些基于细胞的平台通常使用二维基质,而这些基质缺乏支持体内所见三维细胞结构的能力。由于先前已表明单层细胞培养物的功能与体内细胞不同,因此此类体外测试的结果可能无法准确反映体内的细胞反应。因此,确定基于三维细胞的平台中基质结构、细胞结构和细胞功能之间的关系很有意义。为了研究基质结构对胰岛素瘤组织和功能的影响,将胰岛素瘤接种到具有三种不同纤维直径和纤维密度的二维明胶基质和三维纤维状明胶支架上。在培养第3、5和7天评估细胞活力和聚集情况,并在第7天测量基础胰岛素分泌和葡萄糖刺激的胰岛素产生。细小且间距紧密的明胶纤维促进了大的圆形胰岛素瘤簇的形成,而单层组织和粗大纤维则阻止了细胞聚集并降低了葡萄糖刺激的胰岛素产生。综上所述,这些数据表明支架特性可用于控制胰岛素产生细胞的组织和功能,并且可能作为糖尿病药物开发的三维测试平台有用。