Atomic force microscopy combined with human pluripotent stem cell derived cardiomyocytes for biomechanical sensing.

Cardiomyocyte contraction and relaxation are important parameters of cardiac function altered in many heart pathologies. Biosensing of these parameters represents an important tool in drug development and disease modeling. Human embryonic stem cells and especially patient specific induced pluripotent stem cell-derived cardiomyocytes are well established as cardiac disease model.. Here, a live stem cell derived embryoid body (EB) based cardiac cell syncytium served as a biorecognition element coupled to the microcantilever probe from atomic force microscope thus providing reliable micromechanical cellular biosensor suitable for whole-day testing. The biosensor was optimized regarding the type of cantilever, temperature and exchange of media; in combination with standardized protocol, it allowed testing of compounds and conditions affecting the biomechanical properties of EB. The studied effectors included calcium , drugs modulating the catecholaminergic fight-or-flight stress response such as the beta-adrenergic blocker metoprolol and the beta-adrenergic agonist isoproterenol. Arrhythmogenic effects were studied using caffeine. Furthermore, with EBs originating from patient's stem cells, this biosensor can help to characterize heart diseases such as dystrophies.

[1]  Liang Guo,et al.  Estimating the risk of drug-induced proarrhythmia using human induced pluripotent stem cell-derived cardiomyocytes. , 2011, Toxicological sciences : an official journal of the Society of Toxicology.

[2]  P. Samorí.  STM and AFM studies on (bio)molecular systems : unravelling the nanoworld , 2008 .

[3]  W. Hoffmann,et al.  Complication Rate of Right Ventricular Endomyocardial Biopsy via the Femoral Approach: A Retrospective and Prospective Study Analyzing 3048 Diagnostic Procedures Over an 11-Year Period , 2008, Circulation.

[4]  Craig R. Copeland,et al.  Mechanical Coupling Between Myofibroblasts and Cardiomyocytes Slows Electric Conduction in Fibrotic Cell Monolayers , 2011, Circulation.

[5]  A. Katz,et al.  Regulation of Cardiac Contraction and Relaxation , 2000, Circulation.

[6]  P. Doevendans,et al.  Cardiomyocyte differentiation of mouse and human embryonic stem cells * , 2002, Journal of anatomy.

[7]  G. Churchill,et al.  Characterization of human embryonic stem cell lines by the International Stem Cell Initiative , 2007, Nature Biotechnology.

[8]  N. Sreeram,et al.  Force Measurements of Human Embryonic Stem Cell‐Derived Cardiomyocytes in an In Vitro Transplantation Model , 2007, Stem cells.

[9]  L. Allen.  Stem cells. , 2003, The New England journal of medicine.

[10]  S. Frisbee,et al.  Changes in Cardiovascular Health in the United States, 2003–2011 , 2015, Journal of the American Heart Association.

[11]  Chikashi Nakamura,et al.  Mechanical sensing of the penetration of various nanoneedles into a living cell using atomic force microscopy. , 2005, Biosensors & bioelectronics.

[12]  Qin Tu,et al.  Atomic force microscope study of tumor cell membranes following treatment with anti-cancer drugs. , 2009, Biosensors & bioelectronics.

[13]  A. Rauk,et al.  Testing synthetic amyloid-β aggregation inhibitor using single molecule atomic force spectroscopy. , 2014, Biosensors & bioelectronics.

[14]  T. Imamura,et al.  Is the internal jugular vein or femoral vein a better approach site for endomyocardial biopsy in heart transplant recipients? , 2015, International heart journal.

[15]  Manish J. Butte,et al.  Atomic Force Mechanobiology of Pluripotent Stem Cell-Derived Cardiomyocytes , 2012, PloS one.

[16]  Lu Zhang,et al.  In-situ detection of resveratrol inhibition effect on epidermal growth factor receptor of living MCF-7 cells by Atomic Force Microscopy. , 2014, Biosensors & bioelectronics.

[17]  Markéta Bébarová,et al.  Advances in patch clamp technique: towards higher quality and quantity. , 2012, General physiology and biophysics.

[18]  A. Hampl,et al.  Decrease in Abundance of Apurinic/Apyrimidinic Endonuclease Causes Failure of Base Excision Repair in Culture‐Adapted Human Embryonic Stem Cells , 2013, Stem cells.

[19]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[20]  Kenji Yasuda,et al.  On-chip in vitro cell-network pre-clinical cardiac toxicity using spatiotemporal human cardiomyocyte measurement on a chip , 2014, Scientific Reports.

[21]  Karina H. Nakayama,et al.  Multi-cellular interactions sustain long-term contractility of human pluripotent stem cell-derived cardiomyocytes. , 2014, American journal of translational research.

[22]  C. Brooks,et al.  The Possible Role of Calcium in Excitation‐Contraction Coupling of Heart Muscle , 1961, Circulation.

[23]  C. Ripplinger,et al.  Contractile and electrophysiologic characterization of optimized self-organizing engineered heart tissue. , 2012, The Annals of thoracic surgery.

[24]  Surena Vahabi,et al.  Atomic Force Microscopy Application in Biological Research: A Review Study , 2013, Iranian journal of medical sciences.

[25]  Kenji Yasuda,et al.  Dependence of the community effect of cultured cardiomyocytes on the cell network pattern. , 2007, Biochemical and biophysical research communications.

[26]  Wolfgang A. Linke,et al.  Gigantic variety: expression patterns of titin isoforms in striated muscles and consequences for myofibrillar passive stiffness , 2004, Journal of Muscle Research & Cell Motility.

[27]  Y. Yazaki,et al.  Signalling pathways for cardiac hypertrophy. , 1998, Cellular signalling.

[28]  A. Meli,et al.  Human Pluripotent Stem Cell-Derived Cardiomyocytes as Research and Therapeutic Tools , 2014, BioMed research international.

[29]  Michael Regnier,et al.  Substrate stiffness increases twitch power of neonatal cardiomyocytes in correlation with changes in myofibril structure and intracellular calcium. , 2011, Biophysical journal.

[30]  A. Hampl,et al.  Expression and Potential Role of Fibroblast Growth Factor 2 and Its Receptors in Human Embryonic Stem Cells , 2005, Stem cells.

[31]  G. Stienen.  Pathomechanisms in heart failure: the contractile connection , 2014, Journal of Muscle Research and Cell Motility.

[32]  B. Bhushan,et al.  Development of AFM-based techniques to measure mechanical properties of nanoscale structures , 2002 .

[33]  D. Elliott,et al.  The use of agarose microwells for scalable embryoid body formation and cardiac differentiation of human and murine pluripotent stem cells. , 2013, Biomaterials.

[34]  L.A. Fearn,et al.  An optical fiber transducer for single myofibril force measurement , 1993, IEEE Transactions on Biomedical Engineering.

[35]  I. Giaever,et al.  Monitoring fibroblast behavior in tissue culture with an applied electric field. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[36]  W. Bloch,et al.  Contractile properties of early human embryonic stem cell-derived cardiomyocytes: beta-adrenergic stimulation induces positive chronotropy and lusitropy but not inotropy. , 2012, Stem cells and development.

[37]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[38]  M. Sepaniak,et al.  Cantilever transducers as a platform for chemical and biological sensors , 2004 .

[39]  Sangyoon J. Han,et al.  Measuring the contractile forces of human induced pluripotent stem cell-derived cardiomyocytes with arrays of microposts. , 2014, Journal of biomechanical engineering.

[40]  P Hunter,et al.  GENE EXPRESSION OF STRETCH‐ACTIVATED CHANNELS AND MECHANOELECTRIC FEEDBACK IN THE HEART , 2006, Clinical and experimental pharmacology & physiology.

[41]  Guohua Liu,et al.  An aptameric graphene nanosensor for label-free detection of small-molecule biomarkers. , 2015, Biosensors & bioelectronics.

[42]  Yohannes Shiferaw,et al.  Mechanisms Underlying the Formation and Dynamics of Subcellular Calcium Alternans in the Intact Rat Heart , 2009, Circulation research.

[43]  Jan Vrbsky,et al.  Forced aggregation and defined factors allow highly uniform-sized embryoid bodies and functional cardiomyocytes from human embryonic and induced pluripotent stem cells , 2014, Heart and Vessels.

[44]  K. Neelsen,et al.  Mutation frequency dynamics in HPRT locus in culture-adapted human embryonic stem cells and induced pluripotent stem cells correspond to their differentiated counterparts. , 2014, Stem cells and development.

[45]  R. J. Podolsky,et al.  Force measurements in skinned muscle fibres , 1969, The Journal of physiology.

[46]  Shih‐Ann Chen,et al.  Temperature regulates the arrhythmogenic activity of pulmonary vein cardiomyocytes. , 2003, Journal of biomedical science.