StemCellFactory I

Automated Production, Expansion and Differentiation of Induced Puripotent Stem Cells

In the first phase of the project (11/2010 - 04/2014) the StemCellFactory production facility was designed and constructed. Automation, standardization and parallelization of all necessary cell culture steps and a comprehensive quality management were implemented.

Meanwhile, the field of stem cell-based drug discovery has progressed. Assay systems are being developed, which include new very efficient methods for genetic modification and which require mature iPS cell-derived neural and cardiac cells. These new developments and challenges are addressed during the first funding period of the project (06/2014 -12/2015).

In particular, the latest developments in genome editing will be implemented in the fully automated workflow. The integrated quality control by fluorescence-based high-throughput microscopy will be further developed and innovative 3D cell culture methods for maturation of neuronal and cardiac cells in bioreactors will be implemented. 3D organoid cultures of neuronal and cardiac cell populations produce a level of maturity that comes closer to fully functional neurons and cardiomyocytes than the cell preparations so far produced in conventional 2D monolayer cultures.

The production of induced pluripotent stem cells (iPS) cells and the subsequent differentiation into different somatic cell types, such as neurons and cardiomyocytes, require numerous very complex manual processing steps. In addition, iPS cell technology bears an enormous potential in personalized medical therapy. However, the patient-specific approach requires extensive parallelization of all processes that are not feasible with conventional manual cell culture methods.

Objectives of the first project phase were therefore (i) the development and establishment of an automated process for reprogramming of somatic cells, (ii) the design and construction of an integrated automated production prototype and (iii) the development of processes for the generation of iPS cells and iPS cell-based products for the pharmaceutical drug development on an industrial scale and with industrial standards, including all necessary quality controls.

The development and establishment of an integrated fully automated processing chain was accomplished by (i) dissecting all biological processes to reprogram, proliferate and differentiate human somatic cells into machine suitable basic processes and (ii) integrating and optimizing all operations of the processing chain. We used state of the art integration-free and feeder cell-free reprogramming methods for iPS cell generation. Furthermore, innovative solutions for individual processing steps were developed. These approaches took full advantage of the potential of automated optical and mechatronic processes of the StemCellFactory, such as automated high-speed microscopy and automated detection and selection of primary human iPS cell colonies for automated harvest and clonal expansion.

In addition, innovative bioreactor-based processes for the expansion of human iPS cells and their neural and cardiac derivatives were developed. This included a custom-made Microplate Dispenser station, which allows high-speed and high-precision seeding of differentiated cells in high-throughput compatible microtiter plate formats.

The production of iPS cells and iPS cell-based cell products for the pharmaceutical industry requires a high level of quality control and documentation throughout the entire production process to ensure high quality standards of cell products. Therefore a wide range of metrology modules were developed and integrated in the StemCellFactory prototype. In addition, a genome-wide validation of the final cell products is carried out by employing DNA chip technology and methods of systems biology and bioinformatics.

The design and construction of the StemCellFactory prototype was completed and the system is now located at LIFE&BRAIN in Bonn. All automated processes were implemented and validated by a custom-made control software.