Faculty of Medicine, Department of Histology and Embryology, Masaryk University, Brno, Czech Republic.
International Clinical Research Center (ICRC), St. Anne's University Hospital, Brno, Czech Republic.
Biotechnol J. 2024 Aug;19(8):e2400240. doi: 10.1002/biot.202400240.
The development of 3D organoids has provided a valuable tool for studying human tissue and organ development in vitro. Cerebral organoids, in particular, offer a unique platform for investigating neural diseases. However, current methods for generating cerebral organoids suffer from limitations such as labor-intensive protocols and high heterogeneity among organoids. To address these challenges, we present a microfluidic device designed to automate and streamline the formation and differentiation of cerebral organoids. The device utilizes microwells with two different shapes to promote the formation of a single aggregate per well and incorporates continuous medium flow for optimal nutrient exchange. In silico simulations supported the effectiveness of the microfluidic chip in replicating cellular microenvironments. Our results demonstrate that the microfluidic chip enables uniform growth of cerebral organoids, significantly reducing the hands-on time required for maintenance. Importantly, the performance of the microfluidic system is comparable to the standard 96-well plate format even when using half the amount of culture medium, and the resulting organoids exhibit substantially developed neuroepithelial buds and cortical structures. This study highlights the potential of custom-designed microfluidic technology in improving the efficiency of cerebral organoid culture.
3D 类器官的发展为体外研究人类组织和器官发育提供了宝贵的工具。特别是脑类器官，为研究神经疾病提供了独特的平台。然而，目前生成脑类器官的方法存在一些局限性，例如劳动强度大的方案和类器官之间的高度异质性。为了解决这些挑战，我们提出了一种微流控装置，旨在自动化和简化脑类器官的形成和分化。该装置利用具有两种不同形状的微井来促进每个微井中形成单个聚集体，并结合连续介质流以实现最佳的营养交换。计算机模拟支持了微流控芯片在复制细胞微环境方面的有效性。我们的结果表明，微流控芯片能够使脑类器官均匀生长，显著减少维护所需的手工操作时间。重要的是，即使使用一半量的培养基，微流控系统的性能也可与标准的 96 孔板格式相媲美，并且所得的类器官表现出明显发育的神经上皮芽和皮质结构。这项研究强调了定制微流控技术在提高脑类器官培养效率方面的潜力。
