A novel approach to noninvasive monitoring of dissolved carbon dioxide in small-scale cell culture processes

Abstract

Disposable small-scale vessels are commonly used in cell culture studies in academia as well as early stages of bioprocess development. These types of research are crucial for our understanding about cells and bioprocesses as they provide important information regarding different parameters affecting cells. Dissolved carbon dioxide (DCO₂) is one main parameter affecting cell metabolism. It is also an indicator of cell culture well-being. Despite CO₂ being a critical process parameter, there is a lack of appropriate monitoring system for CO₂ in small-scale vessels. Here, we present a membrane-based noninvasive method for measuring DCO₂ in cell culture medium. The idea was achieved by modifying a T-flask and replacing a small area of it with CO₂ permeable silicone membrane. In the proposed method, the concentration of CO₂ dissolved in the cell culture medium is determined by measuring the initial diffusion rate of CO₂ through a silicone membrane attached to the bottom wall of the T-flask. The measurement method was validated previously, and the efficacy of the noninvasive method was evaluated by growing E.coli, Pichia pastoris, and CHO cells in the proposed prototype. The results obtained from this method were verified with other quantitative data obtained from the process such as optical density (OD), cell density, dissolved oxygen (DO) and pH. The results show that the proposed membrane-based method is an effective way for completely noninvasive monitoring of DCO₂ in small-scale cell culture processes. Additional diffusing species such as oxygen could also be measured using the same approach.

Keywords: dissolved CO2 (DCO2); dissolved O2 (DO); mammalian cell culture; noninvasive; rate-based; sensor.