Ensuring defined oxygen levels in stem cell cultivation - a novel approach using DO online monitoringin multiwell plates

摘要

At cultivation of stem cells, defined in-vitroconditions are important. Differentiationlargely depends on the biological environmentand growth conditions. Therefore, monitoringof essential parameters like oxygen and pH isof vital importance and enables systemicoptimization of culture conditions for complexdifferentiation processes.Differentiation of haematopoietic andneuronal stem cells needs exact hypoxicconditions to guarantee reproducible results.The SDR SensorDishR Reader enables for thefirst time non-invasive online monitoring ofdissolved oxygen (DO) and pH in 24-wellmultidishes. Optical sensors for DO or pH arelocated at the bottom of each well and read outthrough the bottom of the multidish by a smalland robust 24-channel reader. This reader canbe placed in an incubator or on a shaker forcontinuous monitoring during the wholeperiod of cultivation.The application of the SensorDish Reader isdemonstrated with examples for cultivation ofhuman embryonic stem cells andhaematopoietic stem/progenitor cells (HSPC).The cells were cultivated directly in theOxoDishR for online oxygen monitoring.Different oxygen levels of the gas phase wereapplied and the dissolved oxygen in themedium was measured for the whole time ofthe cultivation. The influence of opening thencubator, lifting the lid of the multidish, andof media change was also investigated as wellas different cell concentrations.With the SDR SensorDishR Reader, negativeeffects can be detected and resolved in time.Effects which had not been considered beforecan now be obtained and regarded, e.g. toohigh oxygen content in the medium can beavoided by media change under hypoxicincubator atmosphere. The non-invasivemethod of the system guarantees continuousmonitoring during the whole cultivation andreduces the risk of contamination.