(476e) Enabling T-Flasks to Improve Bioprocess Development: A Novel Disposable Cell Culture Train From Cell Thawing to Bench-Scale

During the last five years disposable cell culture technology has become increasingly popular for laboratory scale studies and seed culture generation. In particular, T-flasks are disposable low shear devices commonly used for cell culture in both academia and industry settings. However, even this simple single-use technology lack accurate engineering characterization of critical physical parameters such as gas-liquid mass transfer coefficient (kLa), power input per unit volume (P/V), mixing time as well as velocity characteristics. In this work mass transfer and hydrodynamic studies have been carried out in T-flasks, under static and shaken conditions, using patch-based disposable optical sensors and Particle-Image Velocimetry (PIV), respectively. The accurate measurement of critical parameters like kLa and temporal and spatial variations of shear stresses have provided fundamental insight for the successful scale-up from shaken T-flasks to conventional stirred bioreactors. A number of parameters will be monitored including dissolved oxygen (DO), pH, cell density, product titer and glucose to lactate molar ratio at every stage of the process from cell thawing to bench-scale bioreactor. A control experiment will also be investigated using static T-flasks. This research study may provide novel insight into the type of physiological stress (e.g. respiration rate, glucose to lactate molar ratio) experienced by cells during scale-up, from thawing to bench-scale cell culture devices. From a chemical engineering and regulatory perspective, this study could show that process understanding in T-flasks and improved scale-up strategy can allow integration of small Process Scouting Devices (PSDs) with subsequent larger scale cell culture systems. Ultimately, this work aims at integrating fundamental engineering insight, disposable sensors and cell culture technology to facilitate the future development of a disposable T-flask based cell culture train for clone selection and early seed expansion stages.