DataSheet1_Growth and eGFP Production of CHO-K1 Suspension Cells Cultivated From Single Cell to Laboratory Scale.docx

Scaling down bioproduction processes has become a major driving force for more accelerated and efficient process development over the last decades. Especially expensive and time-consuming processes like the production of biopharmaceuticals with mammalian cell lines benefit clearly from miniaturization, due to higher parallelization and increased insights while at the same time decreasing experimental time and costs. Lately, novel microfluidic methods have been developed, especially microfluidic single-cell cultivation (MSCC) devices have been proved to be valuable to miniaturize the cultivation of mammalian cells. So far, growth characteristics of microfluidic cultivated cell lines were not systematically compared to larger cultivation scales; however, validation of a miniaturization tool against initial cultivation scales is mandatory to prove its applicability for bioprocess development. Here, we systematically investigate growth, morphology, and eGFP production of CHO-K1 cells in different cultivation scales ranging from a microfluidic chip (230 nl) to a shake flask (125 ml) and laboratory-scale stirred tank bioreactor (2.0 L). Our study shows a high comparability regarding specific growth rates, cellular diameters, and eGFP production, which proves the feasibility of MSCC as a miniaturized cultivation tool for mammalian cell culture. In addition, we demonstrate that MSCC provides insights into cellular heterogeneity and single-cell dynamics concerning growth and production behavior which, when occurring in bioproduction processes, might severely affect process robustness.

近年来，生物生产过程的缩小化已成为推动过程开发加速与高效化的主要动力。尤其是那些昂贵且耗时的过程，如哺乳动物细胞系生产的生物制药，从微型化中明显受益，这得益于更高的并行化程度和更深入的洞察力，同时降低了实验时间和成本。最近，开发了新型微流控方法，尤其是微流控单细胞培养（MSCC）设备已被证实对于微型化哺乳细胞培养具有价值。迄今为止，微流控培养的细胞系生长特性尚未与较大培养规模进行系统比较；然而，验证微型化工具与初始培养规模的兼容性对于证明其在生物过程开发中的应用性是必不可少的。在本研究中，我们系统地调查了CHO-K1细胞在不同培养规模下的生长、形态和eGFP生产，这些规模从微流控芯片（230 nl）到摇瓶（125 ml）以及实验室规模搅拌罐生物反应器（2.0 L）不等。我们的研究显示，在特定生长速率、细胞直径和eGFP生产方面具有高度可比性，这证明了MSCC作为微型化哺乳细胞培养工具的可行性。此外，我们还证明了MSCC能够揭示细胞异质性和单细胞动态，尤其是在生长和生产行为方面，这些行为在生物生产过程中发生时可能会严重影响过程的稳健性。