Kaiser Nicholas J, Munarin Fabiola, Coulombe Kareen L K
Center for Biomedical Engineering,, Brown University.
Center for Biomedical Engineering,, Brown University;
J Vis Exp. 2018 May 21(135):57239. doi: 10.3791/57239.
As the field of tissue engineering has continued to mature, there has been increased interest in a wide range of tissue parameters, including tissue shape. Manipulating tissue shape on the micrometer to centimeter scale can direct cell alignment, alter effective mechanical properties, and address limitations related to nutrient diffusion. In addition, the vessel in which a tissue is prepared can impart mechanical constraints on the tissue, resulting in stress fields that can further influence both the cell and matrix structure. Shaped tissues with highly reproducible dimensions also have utility for in vitro assays in which sample dimensions are critical, such as whole tissue mechanical analysis. This manuscript describes an alternative fabrication method utilizing negative master molds prepared from laser etched acrylic: these molds perform well with polydimethylsiloxane (PDMS), permit designs with dimensions on the centimeter scale and feature sizes smaller than 25 µm, and can be rapidly designed and fabricated at a low cost and with minimal expertise. The minimal time and cost requirements allow for laser etched molds to be rapidly iterated upon until an optimal design is determined, and to be easily adapted to suit any assay of interest, including those beyond the field of tissue engineering.
随着组织工程领域不断成熟，人们对包括组织形状在内的各种组织参数的兴趣日益浓厚。在微米到厘米尺度上操控组织形状可以引导细胞排列、改变有效力学性能，并解决与营养物质扩散相关的限制。此外，制备组织的容器会对组织施加机械约束，从而产生应力场，进一步影响细胞和基质结构。具有高度可重复尺寸的成形组织在体外分析中也很有用，比如在全组织力学分析等对样品尺寸要求严格的分析中。本手稿描述了一种利用由激光蚀刻丙烯酸制成的负性母模的替代制造方法：这些模具与聚二甲基硅氧烷（PDMS）配合良好，允许设计厘米级尺寸且特征尺寸小于25微米，并且可以以低成本、凭借最少的专业知识快速设计和制造。对时间和成本的最低要求使得激光蚀刻模具能够快速迭代，直到确定最佳设计，并且能够轻松适应任何感兴趣的分析，包括组织工程领域之外的分析。
