Lin Zuwan, Wang Wenbo, Liu Ren, Li Qiang, Lee Jaeyong, Hirschler Charles, Liu Jia
John A. Paulson School of Engineering and Applied Sciences, Harvard University, Boston, MA, USA.
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Nat Protoc. 2025 Mar 26. doi: 10.1038/s41596-025-01147-7.
Organoids are in vitro miniaturized cellular models of organs that offer opportunities for studying organ development, disease mechanisms and drug screening. Understanding the complex processes governing organoid development and function requires methods suitable for the continuous, long-term monitoring of cell activities (for example, electrophysiological and mechanical activity) at single-cell resolution throughout the entire three-dimensional (3D) structure. Cyborg organoid technology addresses this need by seamlessly integrating stretchable mesh nanoelectronics with tissue-like properties, such as tissue-level flexibility, subcellular feature size and mesh-like networks, into 3D organoids through a 2D-to-3D tissue reconfiguration process during organogenesis. This approach enables longitudinal, tissue-wide, single-cell functional mapping, thereby overcoming the limitations of existing techniques including recording duration, spatial coverage, and the ability to maintain stable contact with the tissue during organoid development. This protocol describes the fabrication and characterization of stretchable mesh nanoelectronics, their electrical performance, their integration with organoids and the acquisition of long-term functional organoid activity requiring multimodal data analysis techniques. Cyborg organoid technology represents a transformative tool for investigating organoid development and function, with potential for improving in vitro disease models, drug screening and personalized medicine. The procedure is suitable for users within a multidisciplinary team with expertise in stem cell biology, tissue engineering, nanoelectronics fabrication, electrophysiology and data science.
类器官是器官的体外微型化细胞模型，为研究器官发育、疾病机制和药物筛选提供了机会。要了解控制类器官发育和功能的复杂过程，需要适合在整个三维（3D）结构中以单细胞分辨率对细胞活动（例如电生理和机械活动）进行连续、长期监测的方法。半机械人器官技术通过在器官发生过程中通过二维到三维组织重构过程，将具有组织样特性（如组织水平的柔韧性、亚细胞特征尺寸和网状网络）的可拉伸网状纳米电子器件无缝集成到3D类器官中，满足了这一需求。这种方法能够进行纵向、全组织、单细胞功能映射，从而克服了现有技术在记录持续时间、空间覆盖范围以及在类器官发育过程中与组织保持稳定接触能力方面的局限性。本方案描述了可拉伸网状纳米电子器件的制造和表征、其电学性能、与类器官的集成以及需要多模态数据分析技术的长期功能性类器官活动的获取。半机械人器官技术是研究类器官发育和功能的变革性工具，具有改善体外疾病模型、药物筛选和个性化医疗的潜力。该程序适用于多学科团队中具有干细胞生物学、组织工程、纳米电子制造、电生理学和数据科学专业知识的用户。
