Skip to main content

Advertisement

SpringerLink
Log in

Characterization of functional ion channels in human cardiac c-kit+ progenitor cells

  • Original Contribution
  • Published:
Basic Research in Cardiology Aims and scope Submit manuscript

Abstract

Cardiac progenitor cells play an important role in cardiac repair and regeneration; however, their cellular biology and electrophysiology are not understood. The present study characterizes the functional ion channels in human cardiac c-kit+ progenitor cells using whole-cell patch voltage-clamp, RT-PCR, and Western blots. We found that several ionic currents were present in human cardiac c-kit+ progenitor cells, including a large-conductance Ca2+-activated K+ current (BKCa) in 86 % of cells, an inwardly rectifying K+ current (I Kir) in 84 % of cells, a transient outward K+ current (I to) in 47 % of cells, a voltage-gated tetrodotoxin-sensitive Na+ current (I Na,TTX) in 61 % of cells. Molecular identities of these ionic currents were determined with RT-PCR and Western-blot analysis. KCa.1.1 (for BKCa), Kir2.1 (for I Kir), Kv4.2 and Kv4.3 (for I to), Nav1.3 and Nav1.6 (for I Na.TTX) were abundantly expressed in human cardiac c-kit+ progenitor cells, which do not resemble cardiomyocytes at all. These results demonstrate for the first time that four types of ionic currents including BKCa, I to, I Kir, and I Na.TTX, are heterogeneously present in human cardiac c-kit+ cells, which may be involved in regulating cellular physiology.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Bearzi C, Rota M, Hosoda T, Tillmanns J, Nascirnbene A, De Angelis A, Yasuzawa-Amano S, Trofimova I, Siggins RW, LeCapitaine N, Cascapera S, Beltrami AP, D’Alessandro DA, Zias E, Quaini F, Urbanek K, Michler RE, Bolli R, Kajstura J, Leri A, Anversa P (2007) Human cardiac stem cells. Proc Natl Acad Sci USA 104:14068–14073. doi:10.1073/pnas.0706760104

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Beltrami AP, Barlucchi L, Torella D, Baker M, Limana F, Chimenti S, Kasahara H, Rota M, Musso E, Urbanek K, Leri A, Kajstura J, Nadal-Ginard B, Anversa P (2003) Adult cardiac stem cells are multipotent and support myocardial regeneration. Cell 114:763–776

    Article  CAS  PubMed  Google Scholar 

  3. Beltrami AP, Urbanek K, Kajstura J, Yan SM, Finato N, Bussani R, Nadal-Ginard B, Silvestri F, Leri A, Beltrami CA, Anversa P (2001) Evidence that human cardiac myocytes divide after myocardial infarction. N Engl J Med 344:1750–1757. doi:10.1056/NEJM200106073442303

    Article  CAS  PubMed  Google Scholar 

  4. Bolli P, Chaudhry HW (2010) Molecular physiology of cardiac regeneration. Ann NY Acad Sci 1211:113–126. doi:10.1111/j.1749-6632.2010.05814.x

    Article  PubMed  Google Scholar 

  5. Bolli R, Chugh AR, D’Amario D, Loughran JH, Stoddard MF, Ikram S, Beache GM, Wagner SG, Leri A, Hosoda T, Sanada F, Elmore JB, Goichberg P, Cappetta D, Solankhi NK, Fahsah I, Rokosh DG, Slaughter MS, Kajstura J, Anversa P (2011) Cardiac stem cells in patients with ischaemic cardiomyopathy (SCIPIO): initial results of a randomised phase 1 trial. Lancet 378:1847–1857. doi:10.1016/S0140-6736(11)61590-0

    Article  PubMed Central  PubMed  Google Scholar 

  6. Bolli R, Tang XL, Sanganalmath SK, Rimoldi O, Mosna F, Abdel-Latif A, Jneid H, Rota M, Leri A, Kajstura J (2013) Intracoronary delivery of autologous cardiac stem cells improves cardiac function in a porcine model of chronic ischemic cardiomyopathy. Circulation 128:122–131. doi:10.1161/CIRCULATIONAHA.112.001075

    Article  CAS  PubMed  Google Scholar 

  7. Bollini S, Smart N, Riley PR (2011) Resident cardiac progenitor cells: at the heart of regeneration. J Mol Cell Cardiol 50:296–303. doi:10.1016/j.yjmcc.2010.07.006

    Article  CAS  PubMed  Google Scholar 

  8. Chugh AR, Beache GM, Loughran JH, Mewton N, Elmore JB, Kajstura J, Pappas P, Tatooles A, Stoddard MF, Lima JA, Slaughter MS, Anversa P, Bolli R (2012) Administration of cardiac stem cells in patients with ischemic cardiomyopathy: the SCIPIO trial: surgical aspects and interim analysis of myocardial function and viability by magnetic resonance. Circulation 126:S54–S64. doi:10.1161/CIRCULATIONAHA.112.092627

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Coiret G, Borowiec AS, Mariot P, Ouadid-Ahidouch H, Matifat F (2007) The antiestrogen tamoxifen activates BK channels and stimulates proliferation of MCF-7 breast cancer cells. Mol Pharmacol 71:843–851. doi:10.1124/mol.106.028290

    Article  CAS  PubMed  Google Scholar 

  10. D’Amario D, Fiorini C, Campbell PM, Goichberg P, Sanada F, Zheng HQ, Hosoda T, Rota M, Connell JM, Gallegos RP, Welt FG, Givertz MM, Mitchell RN, Leri A, Kajstura J, Pfeffer MA, Anversa P (2011) Functionally competent cardiac stem cells can be isolated from endomyocardial biopsies of patients with advanced cardiomyopathies. Circ Res 108:857–861. doi:10.1161/Circresaha.111.241380

    Article  PubMed Central  PubMed  Google Scholar 

  11. Davis DR, Kizana E, Terrovitis J, Barth AS, Zhang YQ, Smith RR, Miake J, Marban E (2010) Isolation and expansion of functionally-competent cardiac progenitor cells directly from heart biopsies. J Mol Cell Cardiol 49:312–321. doi:10.1016/j.yjmcc.2010.02.019

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Deng XL, Lau CP, Lai K, Cheung KF, Lau GK, Li GR (2007) Cell cycle-dependent expression of potassium channels and cell proliferation in rat mesenchymal stem cells from bone marrow. Cell Prolif 40:656–670. doi:10.1111/j.1365-2184.2007.00458.x

    Article  CAS  PubMed  Google Scholar 

  13. Ellison GM, Vicinanza C, Smith AJ, Aquila I, Leone A, Waring CD, Henning BJ, Stirparo GG, Papait R, Scarfo M, Agosti V, Viglietto G, Condorelli G, Indolfi C, Ottolenghi S, Torella D, Nadal-Ginard B (2013) Adult c-kit(pos) cardiac stem cells are necessary and sufficient for functional cardiac regeneration and repair. Cell 154:827–842. doi:10.1016/j.cell.2013.07.039

    Article  CAS  PubMed  Google Scholar 

  14. Ferreira-Martins J, Rondon-Clavo C, Tugal D, Korn JA, Rizzi R, Padin-Iruegas ME, Ottolenghi S, De Angelis A, Urbanek K, Ide-Iwata N, D’Amario D, Hosoda T, Leri A, Kajstura J, Anversa P, Rota M (2009) Spontaneous calcium oscillations regulate human cardiac progenitor cell growth. Circ Res 105:764–774. doi:10.1161/CIRCRESAHA.109.206698

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Fuentes T, Kearns-Jonker M (2013) Endogenous cardiac stem cells for the treatment of heart failure. Stem Cells Cloning 6:1–12. doi:10.2147/SCCAA.S29221

    PubMed Central  PubMed  Google Scholar 

  16. Gaborit N, Le Bouter S, Szuts V, Varro A, Escande D, Nattel S, Demolombe S (2007) Regional and tissue specific transcript signatures of ion channel genes in the non-diseased human heart. J Physiol (Lond) 582:675–693. doi:10.1113/jphysiol.2006.126714

    Article  CAS  Google Scholar 

  17. Han Y, Chen JD, Liu ZM, Zhou Y, Xia JH, Du XL, Jin MW (2010) Functional ion channels in mouse cardiac c-kit(+) cells. Am J Physiol-Cell Physiol 298:C1109–C1117. doi:10.1152/ajpcell.00207.2009

    Article  CAS  PubMed  Google Scholar 

  18. He JQ, Vu DM, Hunt G, Chugh A, Bhatnagar A, Bolli R (2011) Human cardiac stem cells isolated from atrial appendages stably express c-kit. Plos One 6:e27719 doi:10.1371/journal.pone.0027719

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  19. He ML, Liu WJ, Sun HY, Wu W, Liu J, Tse HF, Lau CP, Li GR (2011) Effects of ion channels on proliferation in cultured human cardiac fibroblasts. J Mol Cell Cardiol 51:198–206. doi:10.1016/j.yjmcc.2011.05.008

    Article  CAS  PubMed  Google Scholar 

  20. Heusch G (2011) SCIPIO brings new momentum to cardiac cell therapy. Lancet 378:1827–1828. doi:10.1016/S0140-6736(11)61648-6

    Article  PubMed  Google Scholar 

  21. Hong KU, Li QH, Guo Y, Patton NS, Moktar A, Bhatnagar A, Bolli R (2013) A highly sensitive and accurate method to quantify absolute numbers of c-kit+ cardiac stem cells following transplantation in mice. Basic Res Cardiol 108:346. doi:10.1007/s00395-013-0346-0

    Article  PubMed Central  PubMed  Google Scholar 

  22. Hu H, He ML, Tao R, Sun HY, Hu R, Zang WJ, Yuan BX, Lau CP, Tse HF, Li GR (2009) Characterization of ion channels in human preadipocytes. J Cell Physiol 218:427–435. doi:10.1002/Jcp.21617

    Article  CAS  PubMed  Google Scholar 

  23. Itzhaki-Alfia A, Leor J, Raanani E, Sternik L, Spiegelstein D, Netser S, Holbova R, Pevsner-Fischer M, Lavee J, Barbash IM (2009) Patient characteristics and cell source determine the number of isolated human cardiac progenitor cells. Circulation 120:2559–2566. doi:10.1161/CIRCULATIONAHA.109.849588

    Article  PubMed  Google Scholar 

  24. Jiang P, Rushing SN, Kong CW, Fu J, Lieu DK, Chan CW, Deng W, Li RA (2010) Electrophysiological properties of human induced pluripotent stem cells. Am J Physiol Cell Physiol 298:C486–C495. doi:10.1152/ajpcell.00251.2009

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  25. Kajstura J, Gurusamy N, Ogorek B, Goichberg P, Clavo-Rondon C, Hosoda T, D’Amario D, Bardelli S, Beltrami AP, Cesselli D, Bussani R, del Monte F, Quaini F, Rota M, Beltrami CA, Buchholz BA, Leri A, Anversa P (2010) Myocyte turnover in the aging human heart. Circ Res 107:1374–1386. doi:10.1161/CIRCRESAHA.110.231498

    Article  CAS  PubMed  Google Scholar 

  26. Kuhlmann CRW, Most AK, Li F, Munz BM, Schaefer CA, Walther S, Raedle-Hurst T, Waldecker B, Piper HM, Tillmanns H, Wiecha J (2005) Endothelin-1-induced proliferation of human endothelial cells depends on activation of K+ channels and Ca2+ influx. Acta Physiol Scand 183:161–169

    Article  CAS  PubMed  Google Scholar 

  27. Lagostena L, Avitabile D, De Falco E, Orlandi A, Grassi F, Iachininoto MG, Ragone G, Fucile S, Pompilio G, Eusebi F, Pesce M, Capogrossi MC (2005) Electrophysiological properties of mouse bone marrow c-kit(+) cells co-cultured onto neonatal cardiac myocytes. Cardiovasc Res 66:482–492. doi:10.1016/j.cardiores.2005.01.018

    Article  CAS  PubMed  Google Scholar 

  28. Lei M, Zhang H, Grace AA, Huang CL (2007) SCN5A and sinoatrial node pacemaker function. Cardiovasc Res 74:356–365. doi:10.1016/j.cardiores.2007.01.009

    Article  CAS  PubMed  Google Scholar 

  29. Li GR, Deng XL (2011) Functional ion channels in stem cells. World J Stem Cells 3:19–24. doi:10.4252/wjsc.v3.i3.19

    Article  PubMed Central  PubMed  Google Scholar 

  30. Li GR, Dong MQ (2010) Pharmacology of cardiac potassium channels. Adv Pharmacol 59:93–134. doi:10.1016/S1054-3589(10)59004-5

    Article  PubMed  Google Scholar 

  31. Li GR, Feng J, Wang Z, Fermini B, Nattel S (1996) Adrenergic modulation of ultrarapid delayed rectifier K+ current in human atrial myocytes. Circ Res 78:903–915

    Article  CAS  PubMed  Google Scholar 

  32. Li GR, Feng JL, Yue LX, Carrier M, Nattel S (1996) Evidence for two components of delayed rectifier K+ current in human ventricular myocytes. Circ Res 78:689–696

    Article  CAS  PubMed  Google Scholar 

  33. Li GR, Lau CP, Leung TK, Nattel S (2004) Ionic current abnormalities associated with prolonged action potentials in cardiomyocytes from diseased human right ventricles. Heart Rhythm 1:460–468. doi:10.1016/j.hrthm.2004.06.003

    Article  PubMed  Google Scholar 

  34. Li GR, Sun HY, Chen JB, Zhou Y, Tse HF, Lau CP (2009) Characterization of multiple ion channels in cultured human cardiac fibroblasts. Plos One 4(10):e7307. doi:10.1371/Journal.Pone.0007307

    Article  PubMed Central  PubMed  Google Scholar 

  35. Ma J, Guo L, Fiene SJ, Anson BD, Thomson JA, Kamp TJ, Kolaja KL, Swanson BJ, January CT (2011) High purity human-induced pluripotent stem cell-derived cardiomyocytes: electrophysiological properties of action potentials and ionic currents. Am J Physiol Heart Circ Physiol 301:H2006–H2017. doi:10.1152/ajpheart.00694.2011

    Article  CAS  PubMed  Google Scholar 

  36. Maltsev VA, Wobus AM, Rohwedel J, Bader M, Hescheler J (1994) Cardiomyocytes differentiated in vitro from embryonic stem cells developmentally express cardiac-specific genes and ionic currents. Circ Res 75:233–244

    Article  CAS  PubMed  Google Scholar 

  37. Messina E, De Angelis L, Frati G, Morrone S, Chimenti S, Fiordaliso F, Salio M, Battaglia M, Latronico MV, Coletta M, Vivarelli E, Frati L, Cossu G, Giacomello A (2004) Isolation and expansion of adult cardiac stem cells from human and murine heart. Circ Res 95:911–921. doi:10.1161/01.RES.0000147315.71699.51

    Article  CAS  PubMed  Google Scholar 

  38. Mishra R, Vijayan K, Colletti EJ, Harrington DA, Matthiesen TS, Simpson D, Goh SK, Walker BL, Almeida-Porada G, Wang D, Backer CL, Dudley SC Jr, Wold LE, Kaushal S (2011) Characterization and functionality of cardiac progenitor cells in congenital heart patients. Circulation 123:364–373. doi:10.1161/CIRCULATIONAHA.110.971622

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  39. Nurzynska D, Di Meglio F, Romano V, Miraglia R, Sacco AM, Latino F, Bancone C, Della Corte A, Maiello C, Amarelli C, Montagnani S, Castaldo C (2013) Cardiac primitive cells become committed to a cardiac fate in adult human heart with chronic ischemic disease but fail to acquire mature phenotype: genetic and phenotypic study. Basic Res Cardiol 108:320. doi:10.1007/s00395-012-0320-2

    Article  PubMed  Google Scholar 

  40. Ouadid-Ahidouch H, Roudbaraki M, Delcourt P, Ahidouch A, Joury N, Prevarskaya N (2004) Functional and molecular identification of intermediate-conductance Ca2+-activated K+ channels in breast cancer cells: association with cell cycle progression. Am J Physiol-Cell Physiol 287:C125–C134. doi:10.1152/ajpcell.00488.2003

    Article  CAS  PubMed  Google Scholar 

  41. Pardo LA (2004) Voltage-gated potassium channels in cell proliferation. Physiology (Bethesda) 19:285–292. doi:10.1152/physiol.00011.2004

    Article  CAS  Google Scholar 

  42. Piegari E, De Angelis A, Cappetta D, Russo R, Esposito G, Costantino S, Graiani G, Frati C, Prezioso L, Berrino L, Urbanek K, Quaini F, Rossi F (2013) Doxorubicin induces senescence and impairs function of human cardiac progenitor cells. Basic Res Cardiol 108:334. doi:10.1007/s00395-013-0334-4

    Article  PubMed  Google Scholar 

  43. Sandstedt J, Jonsson M, Kajic K, Sandstedt M, Lindahl A, Dellgren G, Jeppsson A, Asp J (2012) Left atrium of the human adult heart contains a population of side population cells. Basic Res Cardiol 107:255. doi:10.1007/s00395-012-0255-7

    Article  PubMed  Google Scholar 

  44. Smits AM, van Vliet P, Metz CH, Korfage T, Sluijter JP, Doevendans PA, Goumans MJ (2009) Human cardiomyocyte progenitor cells differentiate into functional mature cardiomyocytes: an in vitro model for studying human cardiac physiology and pathophysiology. Nat Protoc 4:232–243. doi:10.1038/nprot.2008.229

    Article  CAS  PubMed  Google Scholar 

  45. Tallini YN, Greene KS, Craven M, Spealman A, Breitbach M, Smith J, Fisher PJ, Steffey M, Hesse M, Doran RM, Woods A, Singh B, Yen A, Fleischmann BK, Kotlikoff MI (2009) c-kit expression identifies cardiovascular precursors in the neonatal heart. Proc Natl Acad Sci USA 106:1808–1813. doi:10.1073/pnas.0808920106

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  46. Tao R, Lau CP, Tse HF, Li GR (2008) Regulation of cell proliferation by intermediate-conductance Ca2+-activated potassium and volume-sensitive chloride channels in mouse mesenchymal stem cells. Am J Physiol-Cell Physiol 295:C1409–C1416. doi:10.1152/ajpcell.00268.2008

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  47. Tillmanns J, Rota M, Hosoda T, Misao Y, Esposito G, Gonzalez A, Vitale S, Parolin C, Yasuzawa-Amano S, Muraski J, De Angelis A, Lecapitaine N, Siggins RW, Loredo M, Bearzi C, Bolli R, Urbanek K, Leri A, Kajstura J, Anversa P (2008) Formation of large coronary arteries by cardiac progenitor cells. Proc Natl Acad Sci USA 105:1668–1673. doi:10.1073/pnas.0706315105

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  48. Wang K, Xue T, Tsang SY, Van Huizen R, Wong CW, Lai KW, Ye Z, Cheng L, Au KW, Zhang J, Li GR, Lau CP, Tse HF, Li RA (2005) Electrophysiological properties of pluripotent human and mouse embryonic stem cells. Stem Cells 23:1526–1534. doi:10.1634/stemcells.2004-0299

    Article  CAS  PubMed  Google Scholar 

  49. Williams AR, Hatzistergos KE, Addicott B, McCall F, Carvalho D, Suncion V, Morales AR, Da Silva J, Sussman MA, Heldman AW, Hare JM (2013) Enhanced effect of combining human cardiac stem cells and bone marrow mesenchymal stem cells to reduce infarct size and to restore cardiac function after myocardial infarction. Circulation 127:213–223. doi:10.1161/CIRCULATIONAHA.112.131110

    Article  PubMed Central  PubMed  Google Scholar 

  50. Yang Y, Li PY, Cheng J, Mao L, Wen J, Tan XQ, Liu ZF, Zeng XR (2013) Function of BKCa channels is reduced in human vascular smooth muscle cells from Han Chinese patients with hypertension. Hypertension 61:519–525. doi:10.1161/HYPERTENSIONAHA.111.00211

    Article  CAS  PubMed  Google Scholar 

  51. Zaruba MM, Soonpaa M, Reuter S, Field LJ (2010) Cardiomyogenic potential of C-kit(+)-expressing cells derived from neonatal and adult mouse hearts. Circulation 121:1992–2000. doi:10.1161/CIRCULATIONAHA.109.909093

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  52. Zhang YH, Sun HY, Chen KH, Du XL, Liu B, Cheng LC, Li X, Jin MW, Li GR (2012) Evidence for functional expression of TRPM7 channels in human atrial myocytes. Basic Res Cardiol 107:282. doi:10.1007/s00395-012-0282-4

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a General Research Fund (771712M) from Research Grant Council of Hong Kong. Ying-Ying Zhang and Hui Che were supported by postgraduate scholarship from the University of Hong Kong.

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China

    Ying-Ying Zhang, Gang Li, Hui Che, Hai-Ying Sun & Gui-Rong Li

  2. Department of Surgery, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China

    Xin Li & Wing-Kuk Au

  3. Xiamen Heart Center, Zhongshan Hospital, Medical School of Xiamen University, Xiamen, Fujian, China

    Gang Li, Guo-Sheng Xiao, Yan Wang & Gui-Rong Li

  4. Department of Physiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, L4-59, Laboratory Block, FMB, 21 Sassoon Road, Pokfulam, Hong Kong, China

    Gui-Rong Li

Authors
  1. Ying-Ying Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hui Che
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hai-Ying Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Wing-Kuk Au
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Guo-Sheng Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Gui-Rong Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yan Wang or Gui-Rong Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, YY., Li, G., Che, H. et al. Characterization of functional ion channels in human cardiac c-kit+ progenitor cells. Basic Res Cardiol 109, 407 (2014). https://doi.org/10.1007/s00395-014-0407-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00395-014-0407-z

Keywords

Search

Navigation