Modular automated microfluidic cell culture platform reduces glycolytic stress in cerebral cortex organoids

Organ-on-a-chip systems combine microfluidics, cell biology, and tissue engineering to culture 3D organ-specific in vitro models that recapitulate the biology and physiology of their in vivo counterparts. Here, we have developed a multiplex platform that automates the culture of individual organoids in isolated microenvironments at user-defined media flow rates. Programmable workflows allow the use of multiple reagent reservoirs that may be applied to direct differentiation, study temporal variables, and grow cultures long term. Novel techniques in polydimethylsiloxane (PDMS) chip fabrication are described here that enable features on the upper and lower planes of a single PDMS substrate. RNA sequencing (RNA-seq) analysis of automated cerebral cortex organoid cultures shows benefits in reducing glycolytic and endoplasmic reticulum stress compared to conventional in vitro cell cultures. Comparative gene expression profiling analysis of RNA-seq data for H9 cerebral cortex organoids cultured in suspension and in automated microfluidics

器官芯片（Organ-on-a-chip）系统结合微流控技术、细胞生物学与组织工程学，用于构建三维器官特异性体外模型，可重现体内对应器官的生物学特性与生理功能。本研究开发了一种多通道平台，可在用户自定义的培养基流速下，于独立微环境中自动化培养单个类器官。该平台搭载可编程工作流，可接入多试剂储库，用于调控细胞定向分化、探究时序变量以及实现长期体外培养。本研究还报道了聚二甲基硅氧烷（polydimethylsiloxane, PDMS）芯片制备的创新工艺，可在单个PDMS基底的上下两面制备微结构。对自动化培养的大脑皮层类器官进行RNA测序（RNA-seq）分析结果显示，相较于传统体外细胞培养体系，该体系可降低糖酵解与内质网应激水平。本研究对悬浮培养与自动化微流控培养的H9大脑皮层类器官的RNA测序数据开展了基因表达谱对比分析。