Advanced Tissue Biofabrication Center, Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, USA.
Biofabrication. 2011 Jun;3(2):025002. doi: 10.1088/1758-5082/3/2/025002. Epub 2011 May 12.
Development of methods for scalable biofabrication of uniformly sized tissue spheroids is essential for tissue spheroid-based bioprinting of large size tissue and organ constructs. The most recent scalable technique for tissue spheroid fabrication employs a micromolded recessed template prepared in a non-adhesive hydrogel, wherein the cells loaded into the template self-assemble into tissue spheroids due to gravitational force. In this study, we present an improved version of this technique. A new mold was designed to enable generation of 61 microrecessions in each well of a 96-well plate. The microrecessions were seeded with cells using an EpMotion 5070 automated pipetting machine. After 48 h of incubation, tissue spheroids formed at the bottom of each microrecession. To assess the quality of constructs generated using this technology, 600 tissue spheroids made by this method were compared with 600 spheroids generated by the conventional hanging drop method. These analyses showed that tissue spheroids fabricated by the micromolded method are more uniform in diameter. Thus, use of micromolded recessions in a non-adhesive hydrogel, combined with automated cell seeding, is a reliable method for scalable robotic fabrication of uniform-sized tissue spheroids.
开发可扩展的生物制造方法来制造大小均匀的组织球体对于基于组织球体的生物打印大尺寸组织和器官构建体至关重要。最近用于组织球体制造的可扩展技术采用在非粘附性水凝胶中制备的微成型凹陷模板，其中由于重力作用，装入模板的细胞自组装成组织球体。在这项研究中，我们提出了该技术的改进版本。设计了一种新的模具，可在 96 孔板的每个孔中产生 61 个微凹陷。使用 EpMotion 5070 自动化移液机在微凹陷中接种细胞。孵育 48 小时后，组织球体在每个微凹陷的底部形成。为了评估使用该技术生成的构建体的质量，将通过该方法生成的 600 个组织球体与通过传统的悬滴法生成的 600 个球体进行比较。这些分析表明，通过微成型方法制造的组织球体在直径上更加均匀。因此，使用非粘附性水凝胶中的微成型凹陷并结合自动化细胞接种是用于可扩展机器人制造均匀大小的组织球体的可靠方法。
