Monitoring pH, temperature and humidity in long-term stem cell culture in CO<inf>2</inf> incubator

Abstract

Cell culture in stem cell research is on the rise, not only for basic research but also for its potential medical and therapeutic applications. Monitoring culture process using sensors throughout the culture helps to optimize culture conditions for optimal growth and maximize yield from the cells. Further, the sensor data gives insight into developing better cell culture systems. Typical stem cell culture platforms, e.g. well plates or flasks, cannot be easily equipped with sensors and impose many challenges in periodic process measurements in a CO <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> incubator. We present an incubator compatible modular measurement system with three sensors, to monitor pH, temperature and humidity continuously throughout the culture. Sensors are assembled around a flow through cuvette for highly sterile non-contact measurements. No sample preparation or sample extraction from the incubator is needed and the measurements are carried out in a closed flow loop without wasting any medium. The modular assembly is novel, reusable and feasible for humid incubator environments. The system has been tested, validated and used in mesenchymal stem cell expansion and differentiation, for periods ranging from two to three weeks. Once the measurement has commenced at the beginning of culture, continuous measurements without sensor recalibration or special manual attention are carried out till the end of the culture. Measurement data clearly show the interplay between measured parameters, indicating a few stress sources present all through the culture. Additionally, it gives an overall picture of behavior of critical control parameters in an incubator and points out the need for bioprocess systems with automatic process monitoring and smart control for maximum yield, optimal growth and maintenance of the cells.