用于强大的人类类器官培养与分析的柱形和灌注板平台
A Pillar and Perfusion Plate Platform for Robust Human Organoid Culture and Analysis.
作者信息
Kang Soo-Yeon, Kimura Masaki, Shrestha Sunil, Lewis Phillip, Lee Sangjoon, Cai Yuqi, Joshi Pranav, Acharya Prabha, Liu Jiafeng, Yang Yong, Sanchez J Guillermo, Ayyagari Sriramya, Alsberg Eben, Wells James M, Takebe Takanori, Lee Moo-Yeal
机构信息
Department of Biomedical Engineering, University of North Texas.
Division of Gastroenterology, Hepatology, and Nutrition, Cincinnati Children's Hospital Medical Center.
出版信息
bioRxiv. 2023 Mar 13:2023.03.11.532210. doi: 10.1101/2023.03.11.532210.
Human organoids have potential to revolutionize disease modeling by providing multicellular architecture and function that are similar to those . This innovative and evolving technology, however, still suffers from assay throughput and reproducibility to enable high-throughput screening (HTS) of compounds due to cumbersome organoid differentiation processes and difficulty in scale-up and quality control. Using organoids for HTS is further challenged by lack of easy-to-use fluidic systems that are compatible with relatively large organoids. Here, we overcome these challenges by engineering "microarray three-dimensional (3D) bioprinting" technology and associated pillar and perfusion plates for human organoid culture and analysis. High-precision, high-throughput stem cell printing and encapsulation techniques were demonstrated on a pillar plate, which was coupled with a complementary deep well plate and a perfusion well plate for static and dynamic organoid culture. Bioprinted cells and spheroids in hydrogels were differentiated into liver and intestine organoids for functional assays. The pillar/perfusion plates are compatible with standard 384-well plates and HTS equipment, and thus may be easily adopted in current drug discovery efforts.
人类类器官通过提供与人体相似的多细胞结构和功能,具有彻底改变疾病建模的潜力。然而,由于类器官分化过程繁琐,以及扩大规模和质量控制方面的困难,这项创新且不断发展的技术在检测通量和可重复性方面仍存在问题,无法实现化合物的高通量筛选(HTS)。使用类器官进行高通量筛选还面临着缺乏与相对较大的类器官兼容的易用流体系统的挑战。在这里,我们通过设计“微阵列三维(3D)生物打印”技术以及用于人类类器官培养和分析的相关柱板和灌注板,克服了这些挑战。在柱板上展示了高精度、高通量的干细胞打印和封装技术,该柱板与用于静态和动态类器官培养的互补深孔板和灌注孔板相结合。水凝胶中的生物打印细胞和球体被分化为肝脏和肠道类器官用于功能检测。柱板/灌注板与标准384孔板和高通量筛选设备兼容,因此可轻松应用于当前的药物发现工作中。