Abstract
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.
Highlights
The number of biotechnologically manufactured products like biopharmaceuticals has increased rapidly over the last decades (Walsh, 2018)
While the bioreactor utilized here consists of a glass vessel, the shake flask is made of plastics, and the applied microfluidic device is a hybrid of glass and PDMS
Environmental control, live cell imaging, and high spatio-temporal resolution make microfluidic single-cell cultivation (MSCC) a highly valuable miniaturization tool as single-cell dynamics under constant cultivation conditions can be analyzed over multiple generations, and intercellular differences in growth behavior or fluorescence-coupled protein expression can be investigated
Summary
The number of biotechnologically manufactured products like biopharmaceuticals has increased rapidly over the last decades (Walsh, 2018). There is a continuing desire for new and more efficient bioprocesses to cover the increasing demand. The development of improved bioprocesses went hand in hand with the technological progress of miniaturization (Hemmerich et al, 2018). Given that mammalian cell culture processes require considerably longer experimental time spans than bacterial processes, and process development is often based on empirical testing of multiple interdependent parameters (Neubauer et al, 2013), especially time reduction and increasing. From Single Cell to Laboratory Scale experimental throughput are highly desirable to enhance process development (Zhang et al, 2010; Rameez et al, 2014). Maximizing the analytical throughput and expanding the degree of parallelization improve process development and cell line or medium design (Betts and Baganz, 2006)
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