Substrate elasticity does not impact DNA methylation changes during differentiation of pluripotent stem cells
Cytotherapy, vol. 26, pp. 1046–1051
Abstract
BACKGROUND AIMS: Substrate elasticity may direct cell-fate decisions of stem cells. However, it is largely unclear how matrix stiffness affects the differentiation of induced pluripotent stem cells (iPSCs) and whether this is also reflected by epigenetic modifications. METHODS: We 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 toward endoderm, mesoderm and ectoderm. RESULTS: Immunofluorescence and gene expression of canonical differentiation markers were 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 did observe epigenetic differences, particularly on mesodermal differentiation. CONCLUSIONS: Stiffness of PDMS substrates did not affect directed differentiation of iPSCs, whereas the moderate epigenetic differences on TCP might also be attributed to other chemical parameters.
Authors 5
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany; Institute for Stem Cell Biology, University Hospital of RWTH Aachen, Aachen, Germany
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
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany; Institute for Stem Cell Biology, University Hospital of RWTH Aachen, Aachen, Germany
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
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany; Institute for Stem Cell Biology, University Hospital of RWTH Aachen, Aachen, Germany
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
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany; Institute for Stem Cell Biology, University Hospital of RWTH Aachen, Aachen, Germany
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
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Universitätsklinikum Aachen · RWTH Aachen University
Affiliation as printed
Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany; Institute for Stem Cell Biology, University Hospital of RWTH Aachen, Aachen, Germany. Electronic address: wwagner@ukaachen.de
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
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