The negligible self-repair potential of the myocardium has led to cell-based tissue engineering approaches to restore heart function. There is more and more consensus that, in addition to cell development, paracrine effects in particular play a pivotal role in the repair of heart tissue. Here, we present two complementary murine P19 and P19CL6 embryonic carcinoma cell-based in vitro test approaches to study the potential of repair cells and the factors secreted by these cells to induce cardiomyogenesis. P19 cells were 3-dimensionally cultured in hanging drops and P19CL6 cells in a monolayer. Both systems, capable of inducible differentiation towards the cardiomyogenic lineage shown by the appearance of beating cells, the expression of connexin 43 and cardiac troponins T and I, were used to test the cardiomyogenesis-inducing potential of human cardiac-derived adherent proliferating (CardAP) cells, which are candidates for heart repair. CardAP cells in coculture as well as CardAP cell-conditioned medium initiated beating in P19 cells, depending on the cell composition and concentration of the medium. CardAP cell-dependent beating was not observed in P19CL6 cultures, but connexin 43 and cardiac troponin formation as well as expression of GATA-binding protein 4 indicated the dose-dependent stimulatory cardiomyogenic effect of human CardAP cells. In summary, in different ways, P19 and P19CL6 cells have shown their capability to detect paracrine effects of human CardAP cells. In a complementary approach, they could be beneficial for determining the stimulatory cardiomyogenic potential of candidate cardiac-repair cells in vitro.

Angoulvant, D., F. Ivanes, R. Ferrera, P.G. Matthews, S. Nataf, M. Ovize (2011) Mesenchymal stem cell conditioned media attenuates in vitro and ex vivo myocardial reperfusion injury. J Heart Lung Transplant 30: 95-102.
Baraniak, P.R., T.C. McDevitt (2010) Stem cell paracrine actions and tissue regeneration. Regener Med 5: 121-143.
Brenner, C., W.M. Franz (2011) The use of stem cells for the repair of cardiac tissue in ischemic heart disease. Expert Rev Med Devices 8: 209-225.
Carvalho, A.B., A.C. de Carvalho (2010) Heart regeneration: past, present and future. World J Cardiol 2: 107-111.
Chen, Y., D.H. Reese (2011) The retinol signaling pathway in mouse pluripotent P19 cells. J Cell Biochem 112: 2865-2872.
Choi, Y.H., T. Saric, B. Nasseri, S. Huhn, S. Van Linthout, R. Hetzer, C. Tschope, C. Stamm (2011) Cardiac cell therapies: the next generation. Cardiovasc Ther 29: 2-16.
Danalache, B.A., J. Gutkowska, M.J. Slusarz, I. Berezowska, M. Jankowski (2010) Oxytocin-Gly-Lys-Arg: a novel cardiomyogenic peptide. PloS One 5: e13643.
Desplantez, T., E. Dupont, N.J. Severs, R. Weingart (2007) Gap junction channels and cardiac impulse propagation. J Membr Biol 218(1-3): 13-28.
Durocher, D., F. Charron, R. Warren, R.J. Schwartz, M. Nemer (1997) The cardiac transcription factors NKX2-5 and GATA-4 are mutual cofactors. EMBO J 16: 5687-5696.
Eschenhagen, T., S. Blankenberg (2013) Personalized therapy in cardiology: biomarkers, pharmacogenetics and therapy of monogenic diseases. Internist 54: 147-152.
Fathi, F., S. Murasawa, S. Hasegawa, T. Asahara, A.J. Kermani, S.J. Mowla (2009) Cardiac differentiation of P19CL6 cells by oxytocin. Int J Cardiol 134: 75-81.
Haag, M., J. Ritterhof, A. Dimura, K. Miteva, S. Van Linthout, C. Tschope, J. Ringe, M. Sittinger (2013) Pro-angiogenic effect of endomyocardial biopsy-derived cells for cardiac differentiation. Curr Tissue Eng 2: 154-159.
Haag, M., S. Van Linthout, S.E. Schroder, U. Freymann, J. Ringe, C. Tschope, M. Sittinger (2010) Endomyocardial biopsy derived adherent proliferating cells - a potential cell source for cardiac tissue engineering. J Cell Biochem 109: 564-575.
Habara-Ohkubo, A. (1996) Differentiation of beating cardiac muscle cells from a derivative of P19 embryonal carcinoma cells. Cell Struct Funct 21: 101-110.
Hu, D.L., F.K. Chen, Y.Q. Liu, Y.H. Sheng, R. Yang, X.Q. Kong, K.J. Cao, H.T. Gu, L.M. Qian (2010) GATA-4 promotes the differentiation of P19 cells into cardiac myocytes. Int J Mol Med 26: 365-372.
Jasmin, D.C. Spray, A.C. Campos de Carvalho, R. Mendez-Otero (2010) Chemical induction of cardiac differentiation in P19 embryonal carcinoma stem cells. Stem Cells Dev 19: 403-412.
Jiang, B., H. Dong, Q. Li, Y. Yu, Z. Zhang, Y. Zhang, G. Wang, Z. Zhang (2013) Differentiation of reprogrammed mouse cardiac fibroblasts into functional cardiomyocytes. Cell Biochem Biophys 66: 309-318.
Kami, D., I. Shiojima, H. Makino, K. Matsumoto, Y. Takahashi, R. Ishii, A.T. Naito, M. Toyoda, H. Saito, M. Watanabe, I. Komuro, A. Umezawa (2008) Gremlin enhances the determined path to cardiomyogenesis. PloS One 3: e2407.
Keiichi, F. (2002) Reprogramming of bone marrow mesenchymal stem cells into cardiomyocytes. C R Biol 325: 1027-1038.
Li, L., H. Gong, H. Yu, X. Liu, Q. Liu, G. Yan, Y. Zhang, H. Lu, Y. Zou, P. Yang (2012a) Knockdown of nucleosome assembly protein 1-like 1 promotes dimethyl sulfoxide-induced differentiation of P19CL6 cells into cardiomyocytes. J Cell Biochem 113: 3788-3796.
Li, T.S., K. Cheng, K. Malliaras, R.R. Smith, Y. Zhang, B. Sun, N. Matsushita, A. Blusztajn, J. Terrovitis, H. Kusuoka, L. Marban, E. Marban (2012b) Direct comparison of different stem cell types and subpopulations reveals superior paracrine potency and myocardial repair efficacy with cardiosphere-derived cells. J Am Coll Cardiol 59: 942-953.
Maltais, S., J.P. Tremblay, L.P. Perrault, H.Q. Ly (2010) The paracrine effect: pivotal mechanism in cell-based cardiac repair. J Cardiovasc Transl Res 3: 652-662.
Mathers, C., J.T. Boerma, D. Ma Fat (2008) The global burden of disease: 2004 update. Geneva, World Health Organization.
McBurney, M.W. (1993) P19 embryonal carcinoma cells. Int J Develop Biol 37: 135-140.
Mirotsou, M., T.M. Jayawardena, J. Schmeckpeper, M. Gnecchi, V.J. Dzau (2011) Paracrine mechanisms of stem cell reparative and regenerative actions in the heart. J Mol Cell Cardiol 50: 280-289.
Miteva, K., M. Haag, J. Peng, K. Savvatis, P.M. Becher, M. Seifert, K. Warstat, D. Westermann, J. Ringe, M. Sittinger, H.P. Schultheiss, C. Tschope, S. Van Linthout (2011) Human cardiac-derived adherent proliferating cells reduce murine acute Coxsackievirus B3-induced myocarditis. PloS One 6: e28513.
Moerkamp, A.T., M.J. Goumans (2012) Cardiac regeneration: stem cells and beyond. Curr Med Chem 19: 5993-6002.
Mordwinkin, N.M., P.W. Burridge, J.C. Wu (2013) A review of human pluripotent stem cell-derived cardiomyocytes for high-throughput drug discovery, cardiotoxicity screening, and publication standards. J Cardiovasc Transl Res 6: 22-30.
Moretti, A., M. Bellin, C.B. Jung, T.M. Thies, Y. Takashima, A. Bernshausen, M. Schiemann, S. Fischer, S. Moosmang, A.G. Smith, J.T. Lam, K.L. Laugwitz (2010) Mouse and human induced pluripotent stem cells as a source for multipotent Isl1+ cardiovascular progenitors. FASEB J 24: 700-711.
Mueller, I., R. Kobayashi, T. Nakajima, M. Ishii, K. Ogawa (2010) Effective and steady differentiation of a clonal derivative of P19CL6 embryonal carcinoma cell line into beating cardiomyocytes. J Biomed Biotechnol 2010: 380561.
Nakanishi, C., M. Yamagishi, K. Yamahara, I. Hagino, H. Mori, Y. Sawa, T. Yagihara, S. Kitamura, N. Nagaya (2008) Activation of cardiac progenitor cells through paracrine effects of mesenchymal stem cells. Biochem Biophys Res Commun 374: 11-16.
Ohtsu, Y., K. Johkura, K. Ito, T. Akashima, K. Asanuma, N. Ogiwara, T. Oka, I. Komuro, K. Sasaki, J. Amano (2005) Stimulation of P19CL6 with multiple reagents induces pulsating particles in vivo. Curr Med Res Opin 21: 795-803.
Ohtsuki, I., S. Morimoto (2008) Troponin: regulatory function and disorders. Biochem Biophys Res Comm 369: 62-73.
Paquin, J., B.A. Danalache, M. Jankowski, S.M. McCann, J. Gutkowska (2002) Oxytocin induces differentiation of P19 embryonic stem cells to cardiomyocytes. Proc Natl Acad Sci USA 99: 9550-9555.
Pfaffl, M.W. (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29: e45.
Ranganath, S.H., O. Levy, M.S. Inamdar, J.M. Karp (2012) Harnessing the mesenchymal stem cell secretome for the treatment of cardiovascular disease. Cell Stem Cell 10: 244-258.
Ringe, J., G.R. Burmester, M. Sittinger (2012) Regenerative medicine in rheumatic disease-progress in tissue engineering. Nat Rev Rheumatol 8: 493-498.
Sadek, H., B. Hannack, E. Choe, J. Wang, S. Latif, M.G. Garry, D.J. Garry, J. Longgood, D.E. Frantz, E.N. Olson, J. Hsieh, J.W. Schneider (2008) Cardiogenic small molecules that enhance myocardial repair by stem cells. Proc Natl Acad Sci USA 105: 6063-6068.
Uchida, S., S. Fuke, T. Tsukahara (2007) Upregulations of GATA4 and oxytocin receptor are important in cardiomyocyte differentiation processes of P19CL6 cells. J Cell Biochem 100: 629-641.
van der Heyden, M.A., L.H. Defize (2003) Twenty-one years of P19 cells: what an embryonal carcinoma cell line taught us about cardiomyocyte differentiation. Cardiovasc Res 58: 292-302.
Verheule, S., M.J. van Kempen, P.H. te Welscher, B.R. Kwak, H.J. Jongsma (1997) Characterization of gap junction channels in adult rabbit atrial and ventricular myocardium. Circ Res 80: 673-681.
Wilton, S., I. Skerjanc (1999) Factors in serum regulate muscle development in P19 cells. In vitro Cell Develop Biol Anim 35: 175-177.
Xu, X.Q., W. Sun (2013) Perspective from the heart: the potential of human pluripotent stem cell-derived cardiomyocytes. J Cell Biochem 114: 39-46.
Yoshida, Y., S. Yamanaka (2011) iPS cells: a source of cardiac regeneration. J Mol Cell Cardiol 50: 327-332.
Zhang, S.S., R.M. Shaw (2012) Multilayered regulation of cardiac ion channels. Biochim Biophys Acta.
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