StemCellFactory II

In the second StemCellFactory project phase (06/2014 - 12/2015) further developments regarding bioreactor-based 3D cell culture methods to generate iPS cell-based neuronal and cardiac derivatives were executed. During the 18-month project phase, a bioreactor-based process for the standardised generation and long-time differentiation (8 to 10 weeks) of brain- and heart tissue-like 3D cultures was developed. Comparitive analysises of gene expressions of neural stem cells, which were differentiated neuronally under unconventional 2D and 3D cell culture conditions, showed that the last gene expression profile is similar to fully matured neurons.

The evolved methods allow the generation of 3D cell models under scalable terms. These models have a vivo-like physiology due to their complexity and maturity. Therefore, they are an attractive cellular resource for desease research and drug development.

To establish the technical and biological bases for automated generation and modification of cellular desease models on the StemCellFactory platform, an automated process flow for a DNA-based CRISPR/Cas genome editing was developed. Besides the implementation and technical validation of required hardware modules on the liquid handling unit, necessary liquid handling processes for the automated genome editing were programmed.

Moreover, the implemented microscope and plate reader were further developed with the objective of generating a continuous and automated quality control of cell cultures during production. This allows a detection of bacterial contamination during the cell production on the StemCellFactory platform. A microscope- and confluence-based method was developed for planning and performing clonal iPS cell passages on the StemCellFactory facility in 24- and 6-well MTPs. It allows the automated expansion of at least ten passages.