The StemCellFactory: A Modular System Integration for Automated Generation and Expansion of Human Induced Pluripotent Stem Cells

Andreas Elanzew; Bastian Nießing; Daniel Langendoerfer; Oliver Rippel; Tobias Piotrowski; Friedrich Schenk; Michael Kulik; Michael Peitz; Yannik Breitkreuz; Sven Jung; Paul Wanek; Laura Stappert; Robert H Schmitt; Simone Haupt; Martin Zenke; Niels König; Oliver Brüstle

doi:10.3389/fbioe.2020.580352

The StemCellFactory: A Modular System Integration for Automated Generation and Expansion of Human Induced Pluripotent Stem Cells

Front Bioeng Biotechnol. 2020 Nov 9:8:580352. doi: 10.3389/fbioe.2020.580352. eCollection 2020.

Authors

Andreas Elanzew^{1

2}, Bastian Nießing³, Daniel Langendoerfer², Oliver Rippel^{2

3}, Tobias Piotrowski³, Friedrich Schenk³, Michael Kulik³, Michael Peitz^{1

4}, Yannik Breitkreuz^{1

2}, Sven Jung³, Paul Wanek^{5

6}, Laura Stappert², Robert H Schmitt^{3

7}, Simone Haupt^{1

2}, Martin Zenke^{5

6}, Niels König³, Oliver Brüstle^{1

2}

Affiliations

¹ Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital Bonn, Bonn, Germany.
² LIFE&BRAIN GmbH, Cellomics Unit, Bonn, Germany.
³ Fraunhofer Institute for Production Technology, Aachen, Germany.
⁴ Cell Programming Core Facility, University of Bonn Medical Faculty, Bonn, Germany.
⁵ Institute for Biomedical Engineering, Cell Biology, Faculty of Medicine, RWTH Aachen University, Aachen, Germany.
⁶ Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
⁷ Laboratory for Machine Tools and Production, RWTH Aachen University, Aachen, Germany.

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.

Keywords: automation; cell culture; cell production; induced pluripotent stem cells; reprogramming.