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Real-Time Monitoring and Control of Soluble Signaling Factors Enables Enhanced Progenitor Cell Outputs from Human Cord Blood Stem Cell Cultures

机译:实时监测和控制可溶性信号因子使人脐带血干细胞培养物中的祖细胞产量提高

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摘要

Monitoring and control of primary cell cultures is challenging as they are heterogenous and dynamically complex systems. Feedback signaling proteins produced from off-target cell populations can accumulate, inhibiting the production of the desired cell populations. Although culture strategies have been developed to reduce feedback inhibition, they are typically optimized for a narrow range of process parameters and do not allow for a dynamically regulated response. Here we describe the development of a microbead-based process control system for the monitoring and control of endogenously produced signaling factors. This system uses quantum dot barcoded microbeads to assay endogenously produced signaling proteins in the culture media, allowing for the dynamic manipulation of protein concentrations. This monitoring system was incorporated into a fed-batch bioreactor to regulate the accumulation of TGF-β1 in an umbilical cord blood cell expansion system. By maintaining the concentration of TGF-β1 below an upper threshold throughout the culture, we demonstrate enhanced ex vivo expansion of hematopoietic progenitor cells at higher input cell densities and over longer culture periods. This study demonstrates the potential of a fully automated and integrated real-time control strategy in stem cell culture systems, and provides a powerful strategy to achieve highly regulated and intensified in vitro cell manufacturing systems. Biotechnol. Bioeng. 2014;111: 1258–1264. © 2013 The Authors Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
机译:监测和控制原代细胞培养物是具有挑战性的,因为它们是异质且动态复杂的系统。从脱靶细胞群产生的反馈信号蛋白会积累,从而抑制所需细胞群的产生。尽管已经开发了减少反馈抑制的培养策略,但通常针对狭窄的过程参数进行了优化,并且不允许动态调节响应。在这里,我们描述了基于微珠的过程控制系统的开发,用于监视和控制内源性产生的信号因子。该系统使用量子点条形码微珠来分析培养基中内源产生的信号蛋白,从而可以动态控制蛋白浓度。将该监测系统并入补料分批生物反应器中,以调节TGF-β1在脐带血细胞扩增系统中的积累。通过在整个培养过程中保持TGF-β1的浓度低于上限,我们证明了造血祖细胞在更高的输入细胞密度和更长的培养周期中的体外扩增增强。这项研究证明了干细胞培养系统中全自动和集成的实时控制策略的潜力,并为实现高度调控和增强的体外细胞生产系统提供了强大的策略。生物技术。生恩2014; 111:1258–1264。 ©2013作者,生物技术和生物工程,由Wiley Periodicals,Inc.出版。

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