Institute for Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada.
Sci Rep. 2020 Apr 24;10(1):6919. doi: 10.1038/s41598-020-62955-3.
To accelerate the cardiac drug discovery pipeline, we set out to develop a platform that would be capable of quantifying tissue-level functions such as contractile force and be amenable to standard multiwell-plate manipulations. We report a 96-well-based array of 3D human pluripotent stem cell (hPSC)-derived cardiac microtissues - termed Cardiac MicroRings (CaMiRi) - in custom 3D-print-molded multiwell plates capable of contractile force measurement. Within each well, two elastomeric microcantilevers are situated above a circumferential ramp. The wells are seeded with cell-laden collagen, which, in response to the gradual slope of the circumferential ramp, self-organizes around tip-gated microcantilevers to form contracting CaMiRi. The contractile force exerted by the CaMiRi is measured and calculated using the deflection of the cantilevers. Platform responses were robust and comparable across wells, and we used it to determine an optimal tissue formulation. We validated the contractile force response of CaMiRi using selected cardiotropic compounds with known effects. Additionally, we developed automated protocols for CaMiRi seeding, image acquisition, and analysis to enable the measurement of contractile force with increased throughput. The unique tissue fabrication properties of the platform, and the consequent effects on tissue function, were demonstrated upon adding hPSC-derived epicardial cells to the system. This platform represents an open-source contractile force screening system useful for drug screening and tissue engineering applications.
为了加速心脏药物研发管道，我们着手开发一个能够量化组织水平功能（如收缩力）并适用于标准多孔板操作的平台。我们报告了一种基于 96 孔的 3D 人多能干细胞（hPSC）衍生的心脏微组织阵列 - 称为心脏微环（CaMiRi） - 在定制的 3D 打印成型多孔板中，可进行收缩力测量。在每个孔中，两个弹性微悬臂梁位于圆周斜坡上方。孔中接种了细胞负载的胶原蛋白，胶原蛋白会响应圆周斜坡的逐渐倾斜，在尖端门控微悬臂梁周围自组织形成收缩的 CaMiRi。CaMiRi 施加的收缩力通过微悬臂梁的挠度进行测量和计算。平台响应在孔之间是稳健且可比的，我们使用它来确定最佳的组织配方。我们使用具有已知效果的选定心脏毒性化合物验证了 CaMiRi 的收缩力反应。此外，我们开发了 CaMiRi 播种、图像采集和分析的自动化方案，以提高收缩力测量的通量。该平台的独特组织制造特性以及对组织功能的相应影响，在向系统中添加 hPSC 衍生的心外膜细胞时得到了证明。该平台代表了一种用于药物筛选和组织工程应用的开源收缩力筛选系统。
