Optoelectronic Data and Code Library

Abstract— Pluripotent cell types retain several characteristics that make them optimal cell source material for applications in drug development, disease modeling, and therapeutic applications. Human induced pluripotent stem cells (hiPSCs) are currently the most accessible cell source material to cultivate and derive cell-based therapeutic solutions at scale. However, a disconnect exists between quality characteristics of phenotype in the pluripotent state, and downstream metrics for efficacy. Bridging this gap is a major challenge to overcome– Given the plasticity of this cell type, environmental conditioning plays a critical role in guiding phenotype. The objective of this work is to present a scale-down, highly parallelizable approach to acquiring real-time data to inform metrics of hiPSC phenotype throughout the biomanufacturing process. Our methods focused on the development of an optoelectronic instrumentation suite capable of measuring pH, dissolved oxygen, and cell density as important surrogates for phenotype in an optimized hiPSC scale-down expansion bioprocess system. We evaluated system performance through control experiments and targeted perturbation of environmental conditions during hiPSC cultivation, with our results showing promising results. We were successful in obtaining continuous, integrated parametric data throughout cultivation and estimating metabolic characteristics of hiPSC phenotype. In conclusion, this system functions as a proof-of-concept translational tool for development of predictive models and monitoring strategies around the elucidation of phenotypic dynamics within hiPSC biomanufacturing. The significance of this work is in its contribution to closing the gap in our understanding of how upstream phenotype intersects with downstream functionality, in pursuit of global optima in hiPSC biomanufacturing for regenerative medicine.

摘要——多能细胞类型具备多种特性，使其成为药物开发、疾病建模和治疗方案应用中的理想细胞来源材料。目前，人诱导多能干细胞（hiPSCs）是可获取的、用于大规模培养和衍生基于细胞的疗法的最便捷的细胞来源材料。然而，在多能状态下的表型质量特性与下游功效指标之间存在脱节。弥合这一差距是克服的主要挑战——鉴于此类细胞的可塑性，环境调控在引导表型方面发挥着至关重要的作用。本研究的目的是提出一种规模缩小、高度可并行化的方法，以获取实时数据，从而为hiPSC表型在整个生物制造过程中的指标提供信息。我们的方法侧重于开发一套光电子仪器，能够测量pH值、溶解氧和细胞密度，这些是优化hiPSC规模缩小扩展生物工艺系统中山PSC表型的重要替代指标。我们通过控制实验和针对hiPSC培养过程中环境条件的针对性扰动来评估系统性能，我们的结果表明了有希望的结果。我们成功地在整个培养过程中获得了连续、集成的参数数据，并估计了hiPSC表型的代谢特性。总之，该系统作为一种概念验证的转化工具，用于开发预测模型和监测策略，旨在阐明hiPSC生物制造中表型动态的机制。本工作的意义在于其对如何理解上游表型与下游功能交叉的认识贡献，追求再生医学中hiPSC生物制造的全球最优解。