Impact of decellularization on porcine myocardium as scaffold for tissue engineered heart tissue

  • Xiaofeng Ye
  • Haozhe Wang
  • Wenhui Gong
  • Shen Li
  • Haiqing Li
  • Zhe WangEmail author
  • Qiang ZhaoEmail author
Tissue Engineering Constructs and Cell Substrates Original Research
Part of the following topical collections:
  1. Tissue Engineering Constructs and Cell Substrates


Decellularized myocardium has been proposed to construct tissue engineered heart tissue, providing the advantage of natural extracellular architecture. Various decellularization protocols have been developed, but the impact of individual decellularization reagent in the protocol remains unclear. The aim of this study is to evaluate the structural impact of three commonly used decellularization reagents on the porcine myocardium. We decellularized porcine heart tissue with trypsin, Triton X-100 or SDS, and analyzed the morphological characteristics of the remaining tissue by SEM, AFM and two-photon LSM. We further recellularized the scaffold with rat myocardial fibroblasts and cardiomyocytes separately. According to the H&E staining and DNA quantification, SDS decellularized more efficiently in comparison to the other two reagents. Moreover, we found distinct surface microarchitecture differences among groups. The changed structure of tissue might result in varied proliferation myocardial fibroblasts and biophysical performance of the engineered heart tissue. This study demonstrated that the microstructure of decellularized porcine heart tissue vary with decellularization agents. Compared to trypsin and Triton X-100, SDS not only decellularized more efficiently but also preserved the biocompatible microstructure of ECM for recellularization.


Sodium Dodecyl Sulfate Heart Tissue Phosphate Buffer Saline Solution Engineer Heart Tissue Decellularization Process 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We are indebted to W. Bian from the Shanghai Institute of Biochemistry and Cell Biology (SIBCB) for help with two-photon LSM and to S. L. Wang from East China University of Science and Technology for help with AFM.

Compliance with ethical standards


The research was partially financially supported by the National Natural Science Foundation of China (Grant Nos. 81000096, 81200093, 31330029 and 81571826).

Supplementary material

Supplementary material 1 (MOV 4307 kb) Representative video of the rCMs seeded heart tissue, which is decellularized with Trypsin

Supplementary material 2 (MOV 4685 kb) Representative video of the rCMs seeded heart tissue, which is decellularized with SDS

Supplementary material 3 (MOV 4908 kb) Representative video of the rCMs seeded heart tissue, which is decellularized with Triton X-100


  1. 1.
    Sakata Y, Shimokawa H. Epidemiology of heart failure in Asia. Circ J. 2013;77:2209–17.CrossRefGoogle Scholar
  2. 2.
    Ott HC, Matthiesen TS, Goh SK, Black LD, Kren SM, Netoff TI, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart. Nat Med. 2008;14:213–21.CrossRefGoogle Scholar
  3. 3.
    Weymann A, Loganathan S, Takahashi H, Schies C, Claus B, Hirschberg K, et al. Development and evaluation of a perfusion decellularization porcine heart model-generation of 3-dimensional myocardial neoscaffolds. Circ J. 2011;75:852–60.CrossRefGoogle Scholar
  4. 4.
    Arenas-Herrera JE, Ko IK, Atala A, Yoo JJ. Decellularization for whole organ bioengineering. Biomed Mater. 2013;2013(8):014106.CrossRefGoogle Scholar
  5. 5.
    Xu H, Xu B, Yang Q, Li X, Ma X, Xia Q, et al. Comparison of decellularization protocols for preparing a decellularized porcine annulus fibrosus scaffold. PLoS One. 2014;9:e86723. doi: 10.1371/journal.pone.0086723.CrossRefGoogle Scholar
  6. 6.
    Patra C, Ricciardi F, Engel FB. The functional properties of nephronectin: an adhesion molecule for cardiac tissue engineering. Biomaterials. 2012;33:4327–35.CrossRefGoogle Scholar
  7. 7.
    Ye X, Hu X, Wang H, Liu J, Zhao Q. Polyelectrolyte multilayer film on decellularized porcine aortic valve can reduce the adhesion of blood cells without affecting the growth of human circulating progenitor cells. Acta Biomater. 2012;8:1057–67.CrossRefGoogle Scholar
  8. 8.
    Dainese L, Guarino A, Burba I, Esposito G, Pompilio G, Polvani G, et al. Heart valve engineering: decellularized aortic homograft seeded with human cardiac stromal cells. J Heart Valve Dis. 2012;21:125–34.Google Scholar
  9. 9.
    Zhou J, Fritze O, Schleicher M, Wendel HP, Schenke-Layland K, Harasztosi C, et al. Impact of heart valve decellularization on 3-D ultrastructure, immunogenicity and thrombogenicity. Biomaterials. 2010;31:2549–54.CrossRefGoogle Scholar
  10. 10.
    Weymann A, Schmack B, Okada T, Soós P, Istók R, Radovits T, et al. Reendothelialization of human heart valve neoscaffolds using umbilical cord-derived endothelial cells. Circ J. 2013;77:207–16.CrossRefGoogle Scholar
  11. 11.
    Ramesh B, Mathapati S, Galla S, Cherian KM, Guhathakurta S. Crosslinked acellular saphenous vein for small-diameter vascular graft. Asian Cardiovasc Thorac Ann. 2013;21:293–302.CrossRefGoogle Scholar
  12. 12.
    Oberwallner B, Brodarac A, Anić P, Sarić T, Wassilew K, Neef K, et al. Human cardiac extracellular matrix supports myocardial lineage commitment of pluripotent stem cells. Eur J Cardiothorac Surg. 2014;. doi: 10.1093/ejcts/ezu163.Google Scholar
  13. 13.
    Cebotari S, Tudorache I, Ciubotaru A, Boethig D, Sarikouch S, Goerler A, et al. Use of fresh decellularized allografts for pulmonary valve replacement may reduce the reoperation rate in children and young adults: early report. Circulation. 2011;124:S115–23.CrossRefGoogle Scholar
  14. 14.
    Remlinger NT, Wearden PD, Gilbert TW. Procedure for decellularization of porcine heart by retrograde coronary perfusion. J Vis Exp. 2012;70:e50059. doi: 10.3791/50059.Google Scholar
  15. 15.
    Akhyari P, Aubin H, Gwanmesia P, Barth M, Hoffmann S, Huelsmann J, et al. The quest for an optimized protocol for whole-heart decellularization: a comparison of three popular and a novel decellularization technique and their diverse effects on crucial extracellular matrix qualities. Tissue Eng Part C. 2011;17:915–26.CrossRefGoogle Scholar
  16. 16.
    Witzenburg C, Raghupathy R, Kren SM, Taylor DA, Barocas VH. Mechanical changes in the rat right ventricle with decellularization. J Biomech. 2012;45:842–9.CrossRefGoogle Scholar
  17. 17.
    Hooks DA, Trew ML, Caldwell BJ, Sands GB, LeGrice IJ, Smaill BH. Laminar arrangement of ventricular myocytes influences electrical behavior of the heart. Circ Res. 2007;101:e103–12. doi: 10.1161/CIRCRESAHA.107.161075.CrossRefGoogle Scholar
  18. 18.
    Engelmayr GC Jr, Cheng M, Bettinger CJ, Borenstein JT, Langer R, Freed LE. Accordion-like honeycombs for tissue engineering of cardiac anisotropy. Nat Mater. 2008;7:1003–10.CrossRefGoogle Scholar
  19. 19.
    Eitan Y, Sarig U, Dahan N, Machluf M. Acellular cardiac extracellular matrix as a scaffold for tissue engineering: in vitro cell support, remodeling, and biocompatibility. Tissue Eng Part C. 2010;16:671–83.CrossRefGoogle Scholar
  20. 20.
    Salick MR, Napiwocki BN, Sha J, Knight GT, Chindhy SA, Kamp TJ, Ashton RS, Crone WC. Micropattern width dependent sarcomere development in human ESC-derived cardiomyocytes. Biomaterials. 2014;35:4454–64.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Cardiac SurgeryRuijin Hospital, Shanghai Jiaotong UniversityShanghaiPeople’s Republic of China

Personalised recommendations