DEVELOPMENT OF AN AUTOMATION SYSTEM OF CULTURE AND LONGITUDINAL EVALUATION OF PATIENT IPS CELL-DERIVED ORGANOIDS (ASCLEPIOS)

患者诱导多能干细胞衍生类器官培养及纵向评估自动化系统的开发(ASCLEPIOS)

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Abstract

BACKGROUND: Misfolding and aggregation of neurodegenerative disease proteins induce synaptic loss and neuronal cell death in specific regions of central nervous system (CNS). Although discoveries in rodent models have led to a better understanding of the mechanisms leading to neurodegeneration, differences between the rodent and human genomes may have profound effects for developing rodent models. For example, binding sites of RNA binding proteins and single nucleotide variants (SNV) associated with neurodegeneration are less conserved in rodent models. METHOD: Human induced pluripotent stem cells (iPSCs) contain donor genetic background and have the potential to generate any type of cells or tissues. Therefore, the iPSC-derived neurons and tissues, called brain organoids, are expected to be useful for establishing CNS disease models and drug discovery. We have generated several brain region-specific organoids from various patient iPS cell lines. AIMS & OBJECTIVES: There are three challenges in drug discovery using iPSCs; (i) reproducibility of experiments, (ii) coordination of differentiation status among different iPSC lines and (iii) adequate cell supply. RESULTS: To overcome these challenges, we are developing an automation system of culture and longitudinal evaluation of patient iPS cell-derived organoids named ASCLEPiOs. ASCLEPiOs includes two robotic arms, a liquid handling system, two automated CO2 incubators and a confocal image cytometer. DISCUSSION & CONCLUSION: ASCLEPiOs will enable us to execute cell culture experiments and image processing analyses to an accurate schedule. We expect that cell-based assays using vulnerable human neuronal cells and organoids produced by ASCLEPiOs will accelerate the discovery of drug candidates including small-molecules and new modalities for neurodegenerative diseases.

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