Towards Self‐Regulated Bioprocessing: A Compact Benchtop Bioreactor System for Monitored and Controlled 3D Cell and Tissue Culture

Abstract

Bioreactors are crucial tools for the manufacturing of living cell‐based tissue engineered products. However, to reach the market successfully, higher degrees of automation, as well as a decreased footprint still need to be reached. In this study, the use of a benchtop bioreactor for in vitro perfusion culture of scaffold‐based tissue engineering constructs is assessed. A low‐footprint benchtop bioreactor system is designed, comprising a single‐use fluidic components and a bioreactor housing. The bioreactor is operated using an in‐house developed program and the culture environment is monitored by specifically designed sensor ports. A gas‐exchange module is incorporated allowing for heat and mass transfers. Titanium‐based scaffolds are seeded with human periosteum‐derived cells and cultured up to 3 weeks. The benchtop bioreactor constructs are compared to benchmark perfusion systems. Live/Dead stainings, DNA quantifications, glucose consumption, and lactate production assays confirm that the constructs cultured in the benchtop bioreactor grew similarly to the benchmark systems. Manual regulation of the system set points enabled efficient alteration of the culture environment in terms of temperature, pH, and dissolved oxygen. This study provides the necessary basis for the development of low‐footprint, automated, benchtop perfusion bioreactors and enables the implementation of active environment control.

Is It Time to Start Transitioning From 2D to 3D Cell Culture?

“…A novel benchtop bioreactor was recently designed and tested in an article by de Bournonville et al (2019). The bioreactor was built to allow the user to control the internal environment as well as use it in a small space outside of clean room environments.…”

Predicting in vitro human mesenchymal stromal cell expansion based on individual donor characteristics using machine learning

“…For hPDCs we have already shown that this applies since process performance substantially increases in bioreactors [46]. In addition, data regarding cell growth could be either monitored online through imaging [47] or other sensors such as oxygen and lactate data [48], that once validated can provide more dense information throughout cell expansion [49] and not only during passaging. Especially with the development of fully controlled and sensorembedded bioreactor systems [50] such approaches could be embedded in control strategies, 'correcting' for donor dependent variation.…”

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…In a recent study, a novel, alternative intensified cell culture perfusion process resulted in a 2-fold volumetric productivity enhancement as compared to a commercially ready, optimized fed-batch process (Gagnon et al, 2019). In another study, a single-use fluidic components-based perfusion bioreactor system was developed and enabled the implementation of active environmental control (Bournonville et al, 2019). In a study by Bertrand et al (2019), the impact of perfusion cultivation on the intracellular physiological state of a CHO cell line was investigated, revealing decreased mAb productivity as well as a transition phase for metabolites and product quality before reaching steadystate conditions.…”