Skip to main content

Advertisement

SpringerLink
Log in

Development of a Method for Isolation of Mature Cardiomyocytes from Human Heart Biopsy Specimens

  • Published:
Bulletin of Experimental Biology and Medicine Aims and scope

To increase the yield of living cells and their survival, studies were carried out to optimize the method for isolating cardiomyocytes from biopsy specimens excised from the right atrial appendages. It was found that creatine, blebbistatin, and taurine are necessary components of the buffer solution during cardiomyocyte isolation, and that composition of the solutions is a more important factor than their oxygenation.

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

Similar content being viewed by others

References

  1. Ackerman MJ, Mohler PJ. Defining a new paradigm for human arrhythmia syndromes: phenotypic manifestations of gene mutations in ion channel- and transporter-associated proteins. Circ. Res. 2010;107(4):457-465. doi: https://doi.org/10.1161/CIRCRESAHA.110.224592

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Blechschmidt S, Haufe V, Benndorf K, Zimmer T. Voltage-gated Na+ channel transcript patterns in the mammalian heart are species-dependent. Prog. Biophys. Mol. Biol. 2008;98(2-3):309-318. doi: https://doi.org/10.1016/j.pbiomolbio.2009.01.009

    Article  CAS  PubMed  Google Scholar 

  3. Zicha S, Moss I, Allen B, Varro A, Papp J, Dumaine R, Antzelevich C, Nattel S. Molecular basis of species-specific expression of repolarizing K+ currents in the heart. Am. J. Physiol. Heart Circ. Physiol. 2003;285(4):H1641-H1649. doi: https://doi.org/10.1152/ajpheart.00346.2003

    Article  CAS  PubMed  Google Scholar 

  4. Oh JG, Kho C, Hajjar RJ, Ishikawa K. Experimental models of cardiac physiology and pathology. Heart Fail. Rev. 2019;24(4):601-615. doi: https://doi.org/10.1007/s10741-019-09769-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Protze SI, Lee JH, Keller GM. Human pluripotent stem cell-derived cardiovascular cells: from developmental biology to therapeutic applications. Cell Stem Cell. 2019;25(3):311-327. doi: https://doi.org/10.1016/j.stem.2019.07.010

    Article  CAS  PubMed  Google Scholar 

  6. Andrysiak K, Stępniewski J, Dulak J. Human-induced pluripotent stem cell-derived cardiomyocytes, 3D cardiac structures, and heart-on-a-chip as tools for drug research. Pflugers Arch. 2021;473(7):1061-1085. doi: https://doi.org/10.1007/s00424-021-02536-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Ronaldson-Bouchard K, Ma SP, Yeager K, Chen T, Song L, Sirabella D, Morikawa K, Teles D, Yazawa M, Vunjak-Novakovic G. Advanced maturation of human cardiac tissue grown from pluripotent stem cells. Nature. 2018;556:239-243. doi: https://doi.org/10.1038/s41586-018-0016-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Karbassi E, Fenix A, Marchiano S, Muraoka N, Nakamura K, Yang X, Murry CE. Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine. Nat. Rev. Cardiol. 2020;17(6):341-359. doi: https://doi.org/10.1038/s41569-019-0331-x

    Article  PubMed  PubMed Central  Google Scholar 

  9. Slotvitsky MM, Tsvelaya VA, Frolova SR, Dement’eva EV, Agladze KI. The study of the functionality of cardiomyocytes obtained from induced pluripotent stem cells for the modeling of cardiac arrhythmias based on long QT syndrome. Vavilov. Zh. Genet. Selekts. 2018;22(2):187-195. Ruddisn. doi: https://doi.org/10.18699/VJ18.346

  10. Jost N, Virág L, Bitay M, Takács J, Lengyel C, Biliczki P, Nagy Z, Bogáts G, Lathrop DA, Papp JG, Varró A. Restricting excessive cardiac action potential and QT prolongation: a vital role for IKs in human ventricular muscle. Circulation. 2005;112(10):1392-1399. doi: https://doi.org/10.1161/CIRCULATIONAHA.105.550111

    Article  PubMed  Google Scholar 

  11. O’Hara T, Virág L, Varró A, Rudy Y. Simulation of the undiseased human cardiac ventricular action potential: model formulation and experimental validation. PLoS Comput. Biol. 2011;7(5):e1002061. doi: https://doi.org/10.1371/journal.pcbi.1002061

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. ten Tusscher KH, Noble D, Noble PJ, Panfilov AV. A model for human ventricular tissue. Am. J. Physiol. Heart Circ. Physiol. 2004;286(4):H1573-H1589. doi: https://doi.org/10.1152/ajpheart.00794.2003

    Article  PubMed  Google Scholar 

  13. Peeters GA, Sanguinetti MC, Eki Y, Konarzewska H, Renlund DG, Karwande SV, Barry WH. Method for isolation of human ventricular myocytes from single endocardial and epicardial biopsies. Am. J. Physiol. 1995;268(N 4, Pt 2):H1757-H1764. doi: https://doi.org/10.1152/ajpheart.1995.268.4.H1757

  14. Bird SD, Doevendans PA, van Rooijen MA, Brutel de la Riviere A, Hassink RJ, Passier R, Mummery CL. The human adult cardiomyocyte phenotype. Cardiovasc. Res. 2003;58(2):423-434. doi: https://doi.org/10.1016/s0008-6363(03)00253-0

  15. Bistola V, Nikolopoulou M, Derventzi A, Kataki A, Sfyras N, Nikou N, Toutouza M, Toutouzas P, Stefanadis C, Konstadoulakis MM. Long-term primary cultures of human adult atrial cardiac myocytes: cell viability, structural properties and BNP secretion in vitro. Int. J. Cardiol. 2008;131(1):113-122. doi: https://doi.org/10.1016/j.ijcard.2007.10.058

    Article  PubMed  Google Scholar 

  16. Voigt N, Pearman CM, Dobrev D, Dibb KM. Methods for isolating atrial cells from large mammals and humans. J. Mol. Cell. Cardiol. 2015;86:187-198. doi: https://doi.org/10.1016/j.yjmcc.2015.07.006

    Article  CAS  PubMed  Google Scholar 

  17. Zhou B, Shi X, Tang X, Zhao Q, Wang L, Yao F, Hou Y, Wang X, Feng W, Wang L, Sun X, Wang L, Hu S. Functional isolation, culture and cryopreservation of adult human primary cardiomyocytes. Signal Transduct. Target Ther. 2022;7(1):254. doi: https://doi.org/10.1038/s41392-022-01044-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Shumakov DV, Agladze KI, Zybin DI, Popov MA, Frolova SR kyzy, Romanova SG. Patent RU No. 2749986. Method for isolation of cardiomyocytes from human heart tissue. Bull. No. 18. Published June 21, 2021.

  19. Shumakov DV, Agladze KI, Zybin DI, Agafonov EG, Popov MA, Romanova SG, Frolova ShR, Slotvitsky MM, Berezhnoy AK, Tsvelaya VA. Status of INA in atrial cardiomyocytes after administration of cardioplegia. Klin. Eksper. Khir. 2022;10(2):26-32. Russian. doi: https://doi.org/10.33029/2308-1198-2022-10-2-26-32

  20. Frolova SR, Gorbunov VS, Shubina NS, Perepukhov AM, Romanova SG, Agladze KI. Stilbene derivative as a photosensitive compound to control the excitability of neonatal rat cardiomyocytes. Biosci. Rep. 2019;39(1):SR20181849. https://doi.org/10.1042/BSR20181849

  21. Podgurskaya AD, Slotvitsky MM, Tsvelaya VA, Frolova SR, Romanova SG, Balashov VA, Agladze KI. Cyclophosphamide arrhythmogenicity testing using human-induced pluripotent stem cell-derived cardiomyocytes. Sci. Rep. 2021;11(1):2336. doi: https://doi.org/10.1038/s41598-020-79085-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Del Franco A, Ambrosio G, Baroncelli L, Pizzorusso T, Barison A, Olivotto I, Recchia FA, Lombardi CM, Metra M, Ferrari Chen YF, Passino C, Emdin M, Vergaro G. Creatine deficiency and heart failure. Heart Fail. Rev. 2022;27(5):1605-1616. doi: https://doi.org/10.1007/s10741-021-10173-y

    Article  CAS  PubMed  Google Scholar 

  23. Balestrino M. Role of creatine in the heart: health and disease. Nutrients. 2021;13(4):1215. doi: https://doi.org/10.3390/nu13041215

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Dou Y, Arlock P, Arner A. Blebbistatin specifically inhibits actin-myosin interaction in mouse cardiac muscle. Am. J. Physiol. Cell Physiol. 2007;293(3):C1148-C1153. doi: https://doi.org/10.1152/ajpcell.00551.2006

    Article  CAS  PubMed  Google Scholar 

  25. Swift LM, Kay MW, Ripplinger CM, Posnack NG. Stop the beat to see the rhythm: excitation-contraction uncoupling in cardiac research. Am. J. Physiol. Heart Circ. Physiol. 2021;321(6):H1005-H1013. doi: https://doi.org/10.1152/ajpheart.00477.2021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Kamkin AG, Kamkina OV, Shim AL, Bilichenko A, Mitrokhin VM, Kazansky VE, Filatova TS, Abramochkin DV, Mladenov MI. The role of activation of two different sGC binding sites by NO-dependent and NO-independent mechanisms in the regulation of SACs in rat ventricular cardiomyocytes. Physiol. Rep. 2022;10(7):e15246. doi: https://doi.org/10.14814/phy2.15246

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Cai B, Mu X, Gong D, Jiang S, Li J, Meng Q, Bai Y, Liu Y, Wang X, Tan X, Yang B, Lu Y. Difference of sodium currents between pediatric and adult human atrial myocytes: evidence for developmental changes of sodium channels. Int. J. Biol. Sci. 2011;7(6):708-714. doi: https://doi.org/10.7150/ijbs.7.708

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Research Laboratory of Molecular and Cellular Diagnostics, M. F. Vladimirsky Moscow Region Research Clinical Institute, Moscow, Russia

    S. G. Kovalenko, Sh. R. Frolova & K. I. Agladze

  2. Laboratory of Experimental and Cellular Medicine, Moscow Institute of Physics and Technology (National Research University), Dolgoprudny, Moscow region, Russia

    S. G. Kovalenko, Sh. R. Frolova, V. K. Kramkova, A. K. Berezovskii & K. I. Agladze

  3. Department of Heart and Vessels Surgery, M. F. Vladimirsky Moscow Region Research Clinical Institute, Moscow, Russia

    M. A. Popov, D. V. Shumakov, D. I. Zybin, E. G. Agafonov & V. V. Dontsov

Authors
  1. S. G. Kovalenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sh. R. Frolova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. K. Kramkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. K. Berezovskii
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. A. Popov
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D. V. Shumakov
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. D. I. Zybin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. E. G. Agafonov
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. V. V. Dontsov
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. K. I. Agladze
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K. I. Agladze.

Additional information

Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 2, pp. 136-143, June, 2023

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kovalenko, S.G., Frolova, S.R., Kramkova, V.K. et al. Development of a Method for Isolation of Mature Cardiomyocytes from Human Heart Biopsy Specimens. Bull Exp Biol Med 175, 585–591 (2023). https://doi.org/10.1007/s10517-023-05907-x

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10517-023-05907-x

Keywords

Search

Navigation