A 3D magnetic tissue stretcher for remote mechanical control of embryonic stem cell differentiation

The ability to create a 3D tissue structure from individual cells and then to stimulate it at will is a major goal for both the biophysics and regenerative medicine communities. Here we show an integrated set of magnetic techniques that meet this challenge using embryonic stem cells (ESCs). We assessed the impact of magnetic nanoparticles internalization on ESCs viability, proliferation, pluripotency and differentiation profiles. We developed magnetic attractors capable of aggregating the cells remotely into a 3D embryoid body. This magnetic approach to embryoid body formation has no discernible impact on ESC differentiation pathways, as compared to the hanging drop method. It is also the base of the final magnetic device, composed of opposing magnetic attractors in order to form embryoid bodies in situ, then stretch them, and mechanically stimulate them at will. These stretched and cyclic purely mechanical stimulations were sufficient to drive ESCs differentiation towards the mesodermal cardiac pathway.The development of embryoid bodies that are responsive to external stimuli is of great interest in tissue engineering. Here, the authors culture embryonic stem cells with magnetic nanoparticles and show that the presence of magnetic fields could affect their aggregation and differentiation.

[1]  Jeff W M Bulte,et al.  Feridex labeling of mesenchymal stem cells inhibits chondrogenesis but not adipogenesis or osteogenesis , 2004, NMR in biomedicine.

[2]  Tejal A Desai,et al.  Programmed synthesis of three-dimensional tissues , 2015, Nature Methods.

[3]  Piotr Walczak,et al.  Tracking stem cells using magnetic nanoparticles. , 2011, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[4]  O. Lee,et al.  Amine‐surface‐modified superparamagnetic iron oxide nanoparticles interfere with differentiation of human mesenchymal stem cells , 2012, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[5]  T. Hyeon,et al.  Iron oxide nanoparticle-mediated development of cellular gap junction crosstalk to improve mesenchymal stem cells' therapeutic efficacy for myocardial infarction. , 2015, ACS nano.

[6]  J. Dobson,et al.  An in vitro model of mesenchymal stem cell targeting using magnetic particle labelling , 2015, Journal of tissue engineering and regenerative medicine.

[7]  G. Vunjak‐Novakovic,et al.  Channelled scaffolds for engineering myocardium with mechanical stimulation , 2012, Journal of tissue engineering and regenerative medicine.

[8]  K. Nakazawa,et al.  Characterization of mouse embryoid bodies cultured on microwell chips with different well sizes. , 2013, Journal of bioscience and bioengineering.

[9]  Eduardo Marbán,et al.  Magnetic Enhancement of Cell Retention, Engraftment, and Functional Benefit after Intracoronary Delivery of Cardiac-Derived Stem Cells in a Rat Model of Ischemia/Reperfusion , 2012, Cell transplantation.

[10]  C. Wilhelm,et al.  Magneto‐Thermal Metrics Can Mirror the Long‐Term Intracellular Fate of Magneto‐Plasmonic Nanohybrids and Reveal the Remarkable Shielding Effect of Gold , 2017 .

[11]  Lu Zhang,et al.  MRI/SPECT/Fluorescent Tri‐Modal Probe for Evaluating the Homing and Therapeutic Efficacy of Transplanted Mesenchymal Stem Cells in a Rat Ischemic Stroke Model , 2015, Advanced functional materials.

[12]  Caterina Minelli,et al.  Substrate stiffness affects early differentiation events in embryonic stem cells. , 2009, European cells & materials.

[13]  J. Werkmeister,et al.  Temporally degradable collagen–mimetic hydrogels tuned to chondrogenesis of human mesenchymal stem cells , 2016, Biomaterials.

[14]  M. Mercola,et al.  Natural and Synthetic Regulators of Embryonic Stem Cell Cardiogenesis , 2009, Pediatric Cardiology.

[15]  Chad A. Cowan,et al.  Interplay of Oct4 with Sox2 and Sox17: a molecular switch from stem cell pluripotency to specifying a cardiac fate , 2009, The Journal of cell biology.

[16]  Michael S Kallos,et al.  Mass Transfer Limitations in Embryoid Bodies during Human Embryonic Stem Cell Differentiation , 2012, Cells Tissues Organs.

[17]  L. Suggs,et al.  Making cardiomyocytes: How mechanical stimulation can influence differentiation of pluripotent stem cells , 2013, Biotechnology progress.

[18]  David A. Weitz,et al.  Physical forces during collective cell migration , 2009 .

[19]  Chan-HyungKim,et al.  Early Expression of Myocardial HIF-1α in Response to Mechanical Stresses , 2002 .

[20]  P. Zandstra,et al.  Reproducible, Ultra High-Throughput Formation of Multicellular Organization from Single Cell Suspension-Derived Human Embryonic Stem Cell Aggregates , 2008, PloS one.

[21]  Chwee Teck Lim,et al.  Epithelial bridges maintain tissue integrity during collective cell migration. , 2014, Nature materials.

[22]  Ali Khademhosseini,et al.  Controlled-size embryoid body formation in concave microwell arrays. , 2010, Biomaterials.

[23]  James A Bankson,et al.  Three-dimensional tissue culture based on magnetic cell levitation. , 2010, Nature nanotechnology.

[24]  Kimiko Yamamoto,et al.  Fluid shear stress induces differentiation of Flk-1-positive embryonic stem cells into vascular endothelial cells in vitro. , 2005, American journal of physiology. Heart and circulatory physiology.

[25]  M. Lutolf,et al.  Substrate elasticity modulates the responsiveness of mesenchymal stem cells to commitment cues. , 2015, Integrative biology : quantitative biosciences from nano to macro.

[26]  A. Arbab,et al.  Labeling of cells with ferumoxides–protamine sulfate complexes does not inhibit function or differentiation capacity of hematopoietic or mesenchymal stem cells , 2005, NMR in biomedicine.

[27]  C. Lien,et al.  A threshold of GATA4 and GATA6 expression is required for cardiovascular development. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Jin Suo,et al.  Magnetic targeting of human mesenchymal stem cells with internalized superparamagnetic iron oxide nanoparticles. , 2013, Small.

[29]  Yan Huang,et al.  Effect of Cyclic Strain on Cardiomyogenic Differentiation of Rat Bone Marrow Derived Mesenchymal Stem Cells , 2012, PloS one.

[30]  Todd C. McDevitt,et al.  Design Principles for Engineering of Tissues from Human Pluripotent Stem Cells , 2016, Current Stem Cell Reports.

[31]  H. Kurosawa.  Methods for inducing embryoid body formation: in vitro differentiation system of embryonic stem cells. , 2007, Journal of bioscience and bioengineering.

[32]  Mehmet Toner,et al.  Microfabrication-based modulation of embryonic stem cell differentiation. , 2007, Lab on a chip.

[33]  F. Mohr,et al.  Cyclic Mechanical Stretch Induces Cardiomyocyte Orientation and Polarization of the Gap Junction Protein Connexin43 , 2010, Circulation research.

[34]  Jong-Wan Park,et al.  Early Expression of Myocardial HIF-1&agr; in Response to Mechanical Stresses: Regulation by Stretch-Activated Channels and the Phosphatidylinositol 3-Kinase Signaling Pathway , 2002, Circulation research.

[35]  G. Hardiman,et al.  Cyclic stretch of embryonic cardiomyocytes increases proliferation, growth, and expression while repressing Tgf-β signaling. , 2015, Journal of molecular and cellular cardiology.

[36]  Ning Wang,et al.  Force via integrins but not E-cadherin decreases Oct3/4 expression in embryonic stem cells. , 2011, Biochemical and biophysical research communications.

[37]  Byung-Soo Kim,et al.  The effect of cyclic strain on embryonic stem cell-derived cardiomyocytes. , 2008, Biomaterials.

[38]  J. Lakins,et al.  Tissue Mechanics Orchestrate Wnt-Dependent Human Embryonic Stem Cell Differentiation. , 2016, Cell stem cell.

[39]  L. Lartigue,et al.  Managing Magnetic Nanoparticle Aggregation and Cellular Uptake: a Precondition for Efficient Stem‐Cell Differentiation and MRI Tracking , 2013, Advanced healthcare materials.

[40]  N. Dehdilani,et al.  Effect of Superparamagnetic Iron Oxide Nanoparticles-Labeling on Mouse Embryonic Stem Cells , 2015, Cell journal.

[41]  M. Asashima,et al.  Directed induction of anterior and posterior primitive streak by Wnt from embryonic stem cells cultured in a chemically defined serum‐free medium , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[42]  M. Poujade,et al.  Velocity fields in a collectively migrating epithelium. , 2010, Biophysical journal.

[43]  H. Honda,et al.  Construction of Functional Cardiovascular Tissues Using Magnetic Nanoparticles , 2013 .

[44]  Tetsuya S. Tanaka,et al.  Generation of organized germ layers from a single mouse embryonic stem cell , 2014, Nature Communications.

[45]  K. Kornev,et al.  Biological magnetic cellular spheroids as building blocks for tissue engineering. , 2014, Acta biomaterialia.

[46]  H. Honda,et al.  iPS cell sheets created by a novel magnetite tissue engineering method for reparative angiogenesis , 2013, Scientific Reports.

[47]  Ronald A. Li,et al.  Effects of iron oxide nanoparticles on cardiac differentiation of embryonic stem cells. , 2009, Biochemical and biophysical research communications.

[48]  Vladimir Mironov,et al.  Organ printing: tissue spheroids as building blocks. , 2009, Biomaterials.

[49]  Deok‐Ho Kim,et al.  Printing three-dimensional tissue analogues with decellularized extracellular matrix bioink , 2014, Nature Communications.

[50]  George Q. Daley,et al.  Biomechanical forces promote embryonic haematopoiesis , 2009, Nature.

[51]  N. Elvassore,et al.  Functional differentiation of human pluripotent stem cells on a chip , 2015, Nature Methods.

[52]  Albert J. Keung,et al.  Soft microenvironments promote the early neurogenic differentiation but not self-renewal of human pluripotent stem cells. , 2012, Integrative biology : quantitative biosciences from nano to macro.

[53]  Y. Yazaki,et al.  Pulsatile stretch activates mitogen-activated protein kinase (MAPK) family members and focal adhesion kinase (p125(FAK)) in cultured rat cardiac myocytes. , 1999, Biochemical and biophysical research communications.

[54]  Jianping Fu,et al.  Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells , 2014, Nature materials.

[55]  Benjamin J Bondow,et al.  Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. , 2008, Developmental biology.

[56]  L. Wang,et al.  Polymorphism in the Alpha Cardiac Muscle Actin 1 Gene Is Associated to Susceptibility to Chronic Inflammatory Cardiomyopathy , 2013, PloS one.

[57]  H. Cheung,et al.  Cyclic compression maintains viability and induces chondrogenesis of human mesenchymal stem cells in fibrin gel scaffolds. , 2009, Stem cells and development.

[58]  Todd C. McDevitt,et al.  Magnetic manipulation and spatial patterning of multi-cellular stem cell aggregates. , 2011, Integrative biology : quantitative biosciences from nano to macro.

[59]  David J. Mooney,et al.  Growth Factors, Matrices, and Forces Combine and Control Stem Cells , 2009, Science.

[60]  E. Marbán,et al.  Magnetic antibody-linked nanomatchmakers for therapeutic cell targeting , 2014, Nature Communications.

[61]  Jeong Ah Kim,et al.  High-throughput generation of spheroids using magnetic nanoparticles for three-dimensional cell culture. , 2013, Biomaterials.

[62]  G. Charras,et al.  Characterizing the mechanics of cultured cell monolayers , 2012, Proceedings of the National Academy of Sciences.

[63]  L. Motte,et al.  Massive Intracellular Biodegradation of Iron Oxide Nanoparticles Evidenced Magnetically at Single-Endosome and Tissue Levels. , 2016, ACS nano.

[64]  Song Li,et al.  Anisotropic mechanosensing by mesenchymal stem cells , 2006, Proceedings of the National Academy of Sciences.

[65]  Joel Voldman,et al.  Fluid shear stress primes mouse embryonic stem cells for differentiation in a self‐renewing environment via heparan sulfate proteoglycans transduction , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[66]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[67]  Ruogang Zhao,et al.  Decoupling Cell and Matrix Mechanics in Engineered Microtissues Using Magnetically Actuated Microcantilevers , 2013, Advanced materials.

[68]  J. Bulte,et al.  Tracking immune cells in vivo using magnetic resonance imaging , 2013, Nature Reviews Immunology.

[69]  Fei Wang,et al.  Material Properties of the Cell Dictate Stress-induced Spreading and Differentiation in Embryonic Stem Cells Growing Evidence Suggests That Physical Microenvironments and Mechanical Stresses, in Addition to Soluble Factors, Help Direct Mesenchymal-stem-cell Fate. However, Biological Responses to a L , 2022 .

[70]  K. McCloskey,et al.  Adhesive forces in embryonic stem cell cultures , 2011, Cell adhesion & migration.

[71]  A. E. El Haj,et al.  Autonomous magnetic labelling of functional mesenchymal stem cells for improved traceability and spatial control in cell therapy applications , 2016, Journal of tissue engineering and regenerative medicine.

[72]  S. Ogawa,et al.  Mechanical stretch activates the JAK/STAT pathway in rat cardiomyocytes. , 1999, Circulation research.