| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Internal Medicine II, University of Lübeck, D-23538 Lübeck;
* Institute of Neurophysiology, University of Cologne, D-50931 Cologne;
Division of Molecular Immunology, Research Center Borstel, D-23845 Borstel;
Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben; and
§ Department of Immunology, University of Lübeck, D-23538 Lübeck, Germany
1Correspondence: Medizinische Klinik II, Medizinische Universität zu Lübeck, Ratzeburger Allee 160, D-23538 Lübeck, Germany. E-mail: franz{at}medinf.mu-luebeck.de
Ischemic disorders of the heart can cause an irreversible loss of cardiomyocytes resulting in a substantial decrease of cardiac output. The therapy of choice is heart transplantation, a technique that is hampered by the low number of donor organs. In the present study, we describe the specific labeling, rapid but gentle purification and characterization of cardiomyocytes derived from mouse pluripotent embryonic stem (ES) cells. To isolate the subpopulation of ventricular-like cardiomyocytes, ES cells were stable transfected with the enhanced green fluorescent protein (EGFP) under transcriptional control of the ventricular-specific 2.1 kb myosin light chain-2v (MLC-2v) promoter and the 0.5 kb enhancer element of the cytomegalovirus (CMVenh.). First fluorescent cells were detected at day 6 + 8 of differentiation within EBs. Four weeks after initiation of differentiation 25% of the cardiomyocyte population displayed fluorescence. Immunohistochemistry revealed the exclusive cardiomyogenic nature of EGFP-positive cells. This was further corroborated by electrophysiological studies where preferentially ventricular phenotypes, but no pacemaker-like cardiomyocytes, were detected among the EGFP-positive population. The enzymatic digestion of EBs, followed by Percoll gradient centrifugation and fluorescence-activated cell sorting, resulted in a 97% pure population of cardiomyocytes. Based on this study, ventricular-like cardiomyocytes can be generated in vitro from EBs and labeled using CMVenh./MLC-2v-driven marker genes facilitating an efficient purification. This method may become an important tool for future cell replacement therapy of ischemic cardiomyopathy especially after the proof of somatic differentiation of human ES cells in vitro.—Müller, M., Fleischmann, B. K., Selbert, S., Ji, G. J., Endl, E., Middeler, G., Mueller, O. J., Schlenke, P., Frese, S., Wobus, A. M., Hescheler, J., Katus, H. A., Franz, W. M. Selection of ventricular-like cardiomyocytes from ES cells in vitro.
Key Words: embryoid body • cardiac • green fluorescent protein • in vitro differentiation
This article has been cited by other articles:
|
|
 |
|
  N. Gassanov, D. Devost, B. Danalache, N. Noiseux, M. Jankowski, H. H. Zingg, and J. Gutkowska Functional Activity of the Carboxyl-Terminally Extended Oxytocin Precursor Peptide During Cardiac Differentiation of Embryonic Stem Cells Stem Cells, January 1, 2008; 26(1): 45 - 54. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Huber, I. Itzhaki, O. Caspi, G. Arbel, M. Tzukerman, A. Gepstein, M. Habib, L. Yankelson, I. Kehat, and L. Gepstein Identification and selection of cardiomyocytes during human embryonic stem cell differentiation FASEB J, August 1, 2007; 21(10): 2551 - 2563. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Gassanov, M. Jankowski, B. Danalache, D. Wang, R. Grygorczyk, U. C. Hoppe, and J. Gutkowska Arginine Vasopressin-mediated Cardiac Differentiation: INSIGHTS INTO THE ROLE OF ITS RECEPTORS AND NITRIC OXIDE SIGNALING J. Biol. Chem., April 13, 2007; 282(15): 11255 - 11265. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Wang, D. L. Haviland, A. R. Burns, E. Zsigmond, and R. A. Wetsel A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cells PNAS, March 13, 2007; 104(11): 4449 - 4454. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Buggisch, B. Ateghang, C. Ruhe, C. Strobel, S. Lange, M. Wartenberg, and H. Sauer Stimulation of ES-cell-derived cardiomyogenesis and neonatal cardiac cell proliferation by reactive oxygen species and NADPH oxidase J. Cell Sci., March 1, 2007; 120(5): 885 - 894. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. W. van Laake, R. Hassink, P. A. Doevendans, and C. Mummery Heart repair and stem cells J. Physiol., December 1, 2006; 577(2): 467 - 478. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Caspi and L. Gepstein Stem cells for myocardial repair Eur. Heart J. Suppl., September 1, 2006; 8(suppl_E): E43 - E54. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Schwanke, S. Wunderlich, M. Reppel, M. E. Winkler, M. Matzkies, S. Groos, J. Itskovitz-Eldor, A. R. Simon, J. Hescheler, A. Haverich, et al. Generation and Characterization of Functional Cardiomyocytes from Rhesus Monkey Embryonic Stem Cells Stem Cells, June 1, 2006; 24(6): 1423 - 1432. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. J. Muller, B. Leuchs, S. T. Pleger, D. Grimm, W.-M. Franz, H. A. Katus, and J. A. Kleinschmidt Improved cardiac gene transfer by transcriptional and transductional targeting of adeno-associated viral vectors Cardiovasc Res, April 1, 2006; 70(1): 70 - 78. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Kouskoff, G. Lacaud, S. Schwantz, H. J. Fehling, and G. Keller Sequential development of hematopoietic and cardiac mesoderm during embryonic stem cell differentiation PNAS, September 13, 2005; 102(37): 13170 - 13175. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. C. Brodie and H. D. Humes Stem Cell Approaches for the Treatment of Renal Failure Pharmacol. Rev., September 1, 2005; 57(3): 299 - 313. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Keller Embryonic stem cell differentiation: emergence of a new era in biology and medicine Genes & Dev., May 15, 2005; 19(10): 1129 - 1155. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Koyanagi, J. Haendeler, C. Badorff, R. P. Brandes, J. Hoffmann, P. Pandur, A. M. Zeiher, M. Kuhl, and S. Dimmeler Non-canonical Wnt Signaling Enhances Differentiation of Human Circulating Progenitor Cells to Cardiomyogenic Cells J. Biol. Chem., April 29, 2005; 280(17): 16838 - 16842. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Wobus and K. R. Boheler Embryonic Stem Cells: Prospects for Developmental Biology and Cell Therapy Physiol Rev, April 1, 2005; 85(2): 635 - 678. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. David, M. Groebner, and W.-M. Franz Magnetic Cell Sorting Purification of Differentiated Embryonic Stem Cells Stably Expressing Truncated Human CD4 as Surface Marker Stem Cells, April 1, 2005; 23(4): 477 - 482. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Hattan, H. Kawaguchi, K. Ando, E. Kuwabara, J. Fujita, M. Murata, M. Suematsu, H. Mori, and K. Fukuda Purified cardiomyocytes from bone marrow mesenchymal stem cells produce stable intracardiac grafts in mice Cardiovasc Res, February 1, 2005; 65(2): 334 - 344. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Xue, H. C. Cho, F. G. Akar, S.-Y. Tsang, S. P. Jones, E. Marban, G. F. Tomaselli, and R. A. Li Functional Integration of Electrically Active Cardiac Derivatives From Genetically Engineered Human Embryonic Stem Cells With Quiescent Recipient Ventricular Cardiomyocytes: Insights Into the Development of Cell-Based Pacemakers Circulation, January 4, 2005; 111(1): 11 - 20. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. C. Davila, G. G. Cezar, M. Thiede, S. Strom, T. Miki, and J. Trosko Use and Application of Stem Cells in Toxicology Toxicol. Sci., June 1, 2004; 79(2): 214 - 223. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Magnusson, C. Rolny, L. Jakobsson, C. Wikner, Y. Wu, D. J. Hicklin, and L. Claesson-Welsh Deregulation of Flk-1/vascular endothelial growth factor receptor-2 in fibroblast growth factor receptor-1-deficient vascular stem cell development J. Cell Sci., March 15, 2004; 117(8): 1513 - 1523. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. White, P. E. Constantin, and W. C. Claycomb Cardiac physiology at the cellular level: use of cultured HL-1 cardiomyocytes for studies of cardiac muscle cell structure and function Am J Physiol Heart Circ Physiol, March 1, 2004; 286(3): H823 - H829. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. J. Muller, M. Lange, H. Rattunde, H.-P. Lorenzen, M. Muller, N. Frey, C. Bittner, W. Simonides, H. A. Katus, and W.-M. Franz Transgenic rat hearts overexpressing SERCA2a show improved contractility under baseline conditions and pressure overload Cardiovasc Res, August 1, 2003; 59(2): 380 - 389. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-Q. He, Y. Ma, Y. Lee, J. A. Thomson, and T. J. Kamp Human Embryonic Stem Cells Develop Into Multiple Types of Cardiac Myocytes: Action Potential Characterization Circ. Res., July 11, 2003; 93(1): 32 - 39. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sachinidis, B. K. Fleischmann, E. Kolossov, M. Wartenberg, H. Sauer, and J. Hescheler Cardiac specific differentiation of mouse embryonic stem cells Cardiovasc Res, May 1, 2003; 58(2): 278 - 291. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. G. Nir, R. David, M. Zaruba, W.-M. Franz, and J. Itskovitz-Eldor Human embryonic stem cells for cardiovascular repair Cardiovasc Res, May 1, 2003; 58(2): 313 - 323. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Dowell, M. Rubart, K. B.S. Pasumarthi, M. H. Soonpaa, and L. J. Field Myocyte and myogenic stem cell transplantation in the heart Cardiovasc Res, May 1, 2003; 58(2): 336 - 350. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Reffelmann and R. A. Kloner Cellular cardiomyoplasty--cardiomyocytes, skeletal myoblasts, or stem cells for regenerating myocardium and treatment of heart failure? Cardiovasc Res, May 1, 2003; 58(2): 358 - 368. [Full Text] [PDF]
|
|
|
|
|
|
M. J. Goldenthal and J. Marin-Garcia Stem cells and cardiac disorders: an appraisal Cardiovasc Res, May 1, 2003; 58(2): 369 - 377. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Johkura, L. Cui, A. Suzuki, R. Teng, A. Kamiyoshi, S. Okamura, S. Kubota, X. Zhao, K. Asanuma, Y. Okouchi, et al. Survival and function of mouse embryonic stem cell-derived cardiomyocytes in ectopic transplants Cardiovasc Res, May 1, 2003; 58(2): 435 - 443. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. I. Rybkin, D. W. Markham, Z. Yan, R. Bassel-Duby, R. S. Williams, and E. N. Olson Conditional Expression of SV40 T-antigen in Mouse Cardiomyocytes Facilitates an Inducible Switch from Proliferation to Differentiation J. Biol. Chem., April 25, 2003; 278(18): 15927 - 15934. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. He, J. Li, E. Bettiol, and M. E. Jaconi Embryonic Stem Cells: New Possible Therapy for Degenerative Diseases That Affect Elderly People J. Gerontol. A Biol. Sci. Med. Sci., March 1, 2003; 58(3): M279 - 287. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Badorff, R. P. Brandes, R. Popp, S. Rupp, C. Urbich, A. Aicher, I. Fleming, R. Busse, A. M. Zeiher, and S. Dimmeler Transdifferentiation of Blood-Derived Human Adult Endothelial Progenitor Cells Into Functionally Active Cardiomyocytes Circulation, February 25, 2003; 107(7): 1024 - 1032. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gepstein Derivation and Potential Applications of Human Embryonic Stem Cells Circ. Res., November 15, 2002; 91(10): 866 - 876. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. M. Zhang, C. Hartzell, M. Narlow, and S. C. Dudley Jr Stem Cell-Derived Cardiomyocytes Demonstrate Arrhythmic Potential Circulation, September 3, 2002; 106(10): 1294 - 1299. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. R. Boheler, J. Czyz, D. Tweedie, H.-T. Yang, S. V. Anisimov, and A. M. Wobus Differentiation of Pluripotent Embryonic Stem Cells Into Cardiomyocytes Circ. Res., August 9, 2002; 91(3): 189 - 201. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
