The StemCellFactory: A Modular System Integration for Automated Generation and Expansion of Human Induced Pluripotent Stem Cells
Frontiers in Bioengineering and Biotechnology, vol. 8, pp. 580352
Abstract
While human induced pluripotent stem cells (hiPSCs) provide novel prospects for disease-modeling, the high phenotypic variability seen across different lines demands usage of large hiPSC cohorts to decipher the impact of individual genetic variants. Thus, a much higher grade of parallelization, and throughput in the production of hiPSCs is needed, which can only be achieved by implementing automated solutions for cell reprogramming, and hiPSC expansion. Here, we describe the StemCellFactory, an automated, modular platform covering the entire process of hiPSC production, ranging from adult human fibroblast expansion, Sendai virus-based reprogramming to automated isolation, and parallel expansion of hiPSC clones. We have developed a feeder-free, Sendai virus-mediated reprogramming protocol suitable for cell culture processing via a robotic liquid handling unit that delivers footprint-free hiPSCs within 3 weeks with state-of-the-art efficiencies. Evolving hiPSC colonies are automatically detected, harvested, and clonally propagated in 24-well plates. In order to ensure high fidelity performance, we have implemented a high-speed microscope for in-process quality control, and image-based confluence measurements for automated dilution ratio calculation. This confluence-based splitting approach enables parallel, and individual expansion of hiPSCs in 24-well plates or scale-up in 6-well plates across at least 10 passages. Automatically expanded hiPSCs exhibit normal growth characteristics, and show sustained expression of the pluripotency associated stem cell marker TRA-1-60 over at least 5 weeks (10 passages). Our set-up enables automated, user-independent expansion of hiPSCs under fully defined conditions, and could be exploited to generate a large number of hiPSC lines for disease modeling, and drug screening at industrial scale, and quality.
Authors 17
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University of Bonn · University Hospital Bonn · Life & Brain (Germany)
Affiliation as printed
Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn, Bonn, Germany
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
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Fraunhofer Institute for Production Technology IPT
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
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Affiliation as printed
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
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Fraunhofer Institute for Production Technology IPT · Life & Brain (Germany)
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
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Fraunhofer Institute for Production Technology IPT
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
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Fraunhofer Institute for Production Technology IPT
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
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Fraunhofer Institute for Production Technology IPT
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
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University of Bonn · University Hospital Bonn
Affiliation as printed
Cell Programming Core Facility, University of Bonn Medical Faculty, Bonn, Germany
Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn, Bonn, Germany
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University of Bonn · University Hospital Bonn · Life & Brain (Germany)
Affiliation as printed
Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn, Bonn, Germany
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
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Fraunhofer Institute for Production Technology IPT
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
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Affiliation as printed
Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany
Institute for Biomedical Engineering, Cell Biology, Faculty of Medicine, RWTH Aachen University, Aachen, Germany
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Affiliation as printed
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
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Robert Heinrich Schmitt Aachen Fraunhofer Institute for Production Technology Laboratory for Machine Tools and Production (WZL)
Fraunhofer Institute for Production Technology IPT · RWTH Aachen University
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
Laboratory for Machine Tools and Production, RWTH Aachen University, Aachen, Germany
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University of Bonn · University Hospital Bonn · Life & Brain (Germany)
Affiliation as printed
Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn, Bonn, Germany
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
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Affiliation as printed
Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany
Institute for Biomedical Engineering, Cell Biology, Faculty of Medicine, RWTH Aachen University, Aachen, Germany
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Fraunhofer Institute for Production Technology IPT
Affiliation as printed
Fraunhofer Institute for Production Technology, Aachen, Germany
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Oliver Brüstle corresponding
University of Bonn · University Hospital Bonn · Life & Brain (Germany)
Affiliation as printed
Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn, Bonn, Germany
LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany
Cited by 47 stored of 49
Cited by patents worldwide 2 (Lens.org)
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