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January 17, 2024 preprint Open access Aachen paper

Substrate elasticity does not impact on DNA methylation changes during differentiation of pluripotent stem cells

bioRxiv (Cold Spring Harbor Laboratory)

DOI: 10.1101/2024.01.16.575833 Full text (OA) OpenAlex

Abstract

Abstract Substrate elasticity may direct cell-fate decisions of stem cells. However, it is largely unclear how matrix stiffness impacts on differentiation of induced pluripotent stem cells (iPSCs) and if this is also reflected by epigenetic modifications. We have therefore cultured iPSCs on tissue culture plastic (TCP) and polydimethylsiloxane (PDMS) with different Young’s modulus (0.2 kPa, 16 kPa, or 64 kPa) to investigate the sequel on growth and differentiation towards endoderm, mesoderm, and ectoderm. Immunofluorescence and gene expression of canonical differentiation markers was hardly affected by the substrates. Notably, when we analyzed DNA methylation profiles of undifferentiated iPSCs or after three-lineage differentiation, we did not see any significant differences on the three different PDMS elasticities. Only when we compared DNA methylation profiles on PDMS-substrates versus TCP, we observed epigenetic differences, particularly upon mesodermal differentiation. Taken together, stiffness of PDMS-substrates did not impact on directed differentiation of iPSCs, whereas the moderate epigenetic differences on TCP might also be attributed to other chemical parameters.

Authors 5

  1. Mohamed H. Elsafi Mabrouk Aachen Institute for Stem Cell Biology

    Universitätsklinikum Aachen · RWTH Aachen University

    Affiliation as printed

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 52074 Aachen, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 5074 Aachen, Germany

  2. Kira Zeevaert Aachen Institute for Stem Cell Biology

    Universitätsklinikum Aachen · RWTH Aachen University

    Affiliation as printed

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 52074 Aachen, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 5074 Aachen, Germany

  3. Ann-Christine Henneke Aachen Institute for Stem Cell Biology

    Universitätsklinikum Aachen · RWTH Aachen University

    Affiliation as printed

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 52074 Aachen, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 5074 Aachen, Germany

  4. Catharina Maaßen Aachen Institute for Stem Cell Biology

    Universitätsklinikum Aachen · RWTH Aachen University

    Affiliation as printed

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 52074 Aachen, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 5074 Aachen, Germany

  5. Wolfgang Wagner corresponding Aachen Institute for Stem Cell Biology

    Universitätsklinikum Aachen · RWTH Aachen University

    Affiliation as printed

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, 52074, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 52074 Aachen, Germany

    Institute for Stem Cell Biology, University Hospital of RWTH Aachen, 5074 Aachen, Germany

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References 30