Schmitz Julian, Hertel Oliver, Yermakov Boris, Noll Thomas, Grünberger Alexander
Multiscale Bioengineering, Faculty of Technology, Bielefeld University, Bielefeld, Germany.
Center for Biotechnology (CeBiTec), Bielefeld University, Bielefeld, Germany.
Front Bioeng Biotechnol. 2021 Oct 15;9:716343. doi: 10.3389/fbioe.2021.716343. eCollection 2021.
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.
在过去几十年中，缩小生物生产工艺规模已成为加速和高效工艺开发的主要驱动力。特别是像使用哺乳动物细胞系生产生物制药这样昂贵且耗时的工艺，由于更高的并行性和更多的深入了解，同时减少了实验时间和成本，显然从小型化中受益。最近，新型微流控方法得到了发展，特别是微流控单细胞培养（MSCC）装置已被证明对哺乳动物细胞培养的小型化很有价值。到目前为止，微流控培养细胞系的生长特性尚未与更大规模的培养进行系统比较；然而，针对初始培养规模验证小型化工具对于证明其在生物工艺开发中的适用性是必不可少的。在此，我们系统地研究了CHO-K1细胞在从微流控芯片（230微升）到摇瓶（125毫升）以及实验室规模搅拌罐生物反应器（2.0升）等不同培养规模下的生长、形态和eGFP产生情况。我们的研究表明，在比生长速率、细胞直径和eGFP产生方面具有高度可比性，这证明了MSCC作为哺乳动物细胞培养小型化工具的可行性。此外，我们证明MSCC能够深入了解细胞异质性以及关于生长和生产行为的单细胞动态，而这些在生物生产过程中出现时可能会严重影响工艺稳健性。
