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Application note: Production of human induced pluripotent stem cell-derived cortical neurospheres in the DASbox® Mini Bioreactor System

Posted: 29 November 2018 | | No comments yet

There is substantial interest in the scientific community in using human induced pluripotent stem cell (hiPSC)-derived neural cells to study basic mechanisms of brain development, neuronal function, and drug-induced effects. In vitro-generated brain organoids are promising models, because they show a remarkable degree of maturation and can be kept in culture for up to 20 months.

In many established protocols for organoid production, cells are embedded in extracellular matrix components and cultivated under static conditions in plates or dishes. This procedure requires a considerable amount of manual lab work, which limits its scalability. In addition, there is a high degree of inter-organoid heterogeneity caused by stochastic effects during the initial formation of the organoids. These factors make current organoid technologies unsuitable for the development of standardised models or assays.

Cultivation in stirred-tank bioreactors allows the tight regulation of process parameters and therefore improves the homogeneity of neural spheroid cultures. It also allows scale-up to produce sufficient cell numbers for novel pharmacological testing devices such as Multi-Organ-Chip systems. Here, researchers at TissUse GmbH demonstrate the potential of stirred-tank bioreactors for the generation of hiPSC-derived neurospheres. They obtained 2 x 108 cells in a single DASbox Mini Bioreactor. Results from three independent bioprocesses demonstrate the reproducibility of the neural differentiation protocol.

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