The spatial self-organization within pluripotent stem cell colonies is continued in detaching aggregates
Biomaterials, vol. 282, pp. 121389
Abstract
Colonies of induced pluripotent stem cells (iPSCs) reveal aspects of self-organization even under culture conditions that maintain pluripotency. To investigate the dynamics of this process under spatial confinement, we used either polydimethylsiloxane (PDMS) pillars or micro-contact printing of vitronectin. There was a progressive upregulation of OCT4, E-cadherin, and NANOG within 70 μm from the outer rim of iPSC colonies. Single-cell RNA-sequencing and spatial reconstruction of gene expression demonstrated that OCT4high subsets, residing at the edge of the colony, have pronounced up-regulation of the TGF-β pathway, particularly of NODAL and its inhibitor LEFTY. Interestingly, after 5–7 days, iPSC colonies detached spontaneously from micro-contact printed substrates to form 3D aggregates. This new method allowed generation of embryoid bodies (EBs) of controlled size without enzymatic or mechanical treatment. Within the early 3D aggregates, radial organization and differential gene expression continued in analogy to the changes observed during self-organization of iPSC colonies. Early self-detached aggregates revealed up-regulated germline-specific gene expression patterns as compared to conventional EBs. However, there were no marked differences after further directed differentiation toward hematopoietic, mesenchymal, and neuronal lineages. Our results provide further insight into the gradual self-organization within iPSC colonies and at their transition into EBs.
Authors 18
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany. Electronic address: mmabrouk@ukaachen.de
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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St. Anna Children's Cancer Research Institute
Affiliation as printed
St. Anna Children's Cancer Research Institute (CCRI), Zimmermannplatz 10, 1090, Vienna, Austria
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Affiliation as printed
Institute of Pathology, RWTH Aachen University Medical School, Pauwelsstraße 30, 52074, Aachen, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Department of Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Faculty of Medicine, RWTH Aachen University, Aachen, 52074, Germany
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Affiliation as printed
Institute for Computational Genomics, RWTH Aachen University Medical School, Pauwelsstraße 19, 52074, Aachen, Germany
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Affiliation as printed
Institute for Computational Genomics, RWTH Aachen University Medical School, Pauwelsstraße 19, 52074, Aachen, Germany
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Affiliation as printed
Institute of Physiology, RWTH Aachen University Medical School, Pauwelsstraße 30, 52074, Aachen, Germany
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Vivek Pachauri Aachen Institute of Materials in Electrical Engineering 1 Chair of Micro- and Nanosystems
Affiliation as printed
Institute of Materials in Electrical Engineering 1 and Chair of Micro-and Nanosystems, RWTH Aachen University, Aachen, 52074, Germany
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Uwe Schnakenberg Aachen Institute of Materials in Electrical Engineering 1 Chair of Micro- and Nanosystems
Affiliation as printed
Institute of Materials in Electrical Engineering 1 and Chair of Micro-and Nanosystems, RWTH Aachen University, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
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Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany; Institute for Biomedical Engineering - Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Helmholtz-Institute for Biomedical Engineering, Stem Cell Biology and Cellular Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany
Institute for Biomedical Engineering – Cell Biology, RWTH Aachen University Medical School, Aachen, 52074, Germany
Cited by 25 stored of 26
Cited by patents worldwide 2 (Lens.org)
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METHOD OF QUALITATIVE CONTROL OF STEM CELLSWO2024056635A1 2024-03-21 Pending Aachen patent
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Verfahren zur qualitativen Kontrolle von StammzellenDE102023105548A1 2024-03-14 Pending Aachen patent