Skip to main content

Advertisement

SpringerLink
Log in

Design of a Miniature Tissue Culture System to Culture Mouse Heart Valves

  • Published:
Annals of Biomedical Engineering Aims and scope Submit manuscript

Abstract

Valvular heart disease is a leading cause of morbidity and mortality in adults but little is known about the underlying etiology. A better understanding of the genetic and hemodynamic mechanisms involved in growth and remodeling of heart valves during physiological and pathological conditions is needed for a better understanding of valvular heart disease. Here, we report the design of a miniature tissue culture system (MTCS) that allows the culture of mitral valves from perinatal to adult mice. The design of the MTCS is novel in that fine positioning and cannulation can be conducted with hearts of different sizes (perinatal to adult). Perfusion of the heart and hence, culture of the mitral valve in its natural position, occurs in a hydraulically sealed culture bath environment. Using the MTCS, we successfully cultured the mitral valve of adult mouse hearts for 3 days. Histological analysis indicated that the cultured valves remained viable and their extracellular matrix organization was similar to age-matched native valves. Gene expression could also be modified in cultured valves by perfusion with medium containing beta-galactosidase-expressing adenovirus. Thus, the MTCS is a new tool to study the genetic and hemodynamic mechanisms underlying the three-dimensional organization of the heart valves, which could provide insights in the pathology of valvular heart disease and be used in animal models for the development of tissue-engineered heart valves.

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

FIGURE 1
FIGURE 2
FIGURE 3
FIGURE 4
FIGURE 5
FIGURE 6
FIGURE 7

Abbreviations

MTCS:

Miniature tissue culture system

E:

Embryonic day

ETO:

Ethylene oxide

IVS:

Interventricular septum

MiVa:

Mitral valve

LV:

Left ventricle

References

  1. Armstrong, E. J., and J. Bischoff. Heart valve development: endothelial cell signaling and differentiation. Circ. Res. 95:459–470, 2004.

    Article  PubMed  CAS  Google Scholar 

  2. Barnett, J. V., and J. S. Desgrosellier. Early events in valvulogenesis: a signaling perspective. Birth Defects Res. C Embryo Today 69:58–72, 2003.

    Article  PubMed  CAS  Google Scholar 

  3. Barron, V., et al. Bioreactors for cardiovascular cell and tissue growth: a review. Ann. Biomed. Eng. 31:1017–1030, 2003.

    Article  PubMed  CAS  Google Scholar 

  4. Bernanke, D. H., and R. R. Markwald. Migratory behavior of cardiac cushion tissue cells in a collagen-lattice culture system. Dev. Biol. 91:235–245, 1982.

    Article  PubMed  CAS  Google Scholar 

  5. Butcher, J. T., and R. M. Nerem. Valvular endothelial cells and the mechanoregulation of valvular pathology. Philos. Trans. R. Soc. Lond. B Biol. Sci. 362:1445–1457, 2007.

    Article  PubMed  CAS  Google Scholar 

  6. Camenisch, T. D., et al. Temporal and distinct TGFbeta ligand requirements during mouse and avian endocardial cushion morphogenesis. Dev. Biol. 248:170–181, 2002.

    Article  PubMed  CAS  Google Scholar 

  7. Devereux, R. B., et al. Inheritance of mitral valve prolapse: effect of age and sex on gene expression. Ann. Intern. Med. 97:826–832, 1982.

    PubMed  CAS  Google Scholar 

  8. Engelmayr, Jr., G. C., et al. A novel bioreactor for the dynamic flexural stimulation of tissue engineered heart valve biomaterials. Biomaterials 24:2523–2532, 2003.

    Article  PubMed  CAS  Google Scholar 

  9. Gaussin, V., et al. Endocardial cushion and myocardial defects after cardiac myocyte-specific conditional deletion of the bone morphogenetic protein receptor ALK3. Proc. Natl Acad. Sci. USA 99:2878–2883, 2002.

    Article  PubMed  CAS  Google Scholar 

  10. Hove, J. R., et al. Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis. Nature 421:172–177, 2003.

    Article  PubMed  CAS  Google Scholar 

  11. Kruithof, B. P., S. A. Krawitz, and V. Gaussin. Atrioventricular valve development during late embryonic and postnatal stages involves condensation and extracellular matrix remodeling. Dev. Biol. 302:208–217, 2007.

    Article  PubMed  CAS  Google Scholar 

  12. Markwald, R., et al. Epithelial-mesenchymal transformations in early avian heart development. Acta Anat. 156:173–186, 1996.

    Article  PubMed  CAS  Google Scholar 

  13. Mizuguchi, T., and N. Matsumoto. Recent progress in genetics of Marfan syndrome and Marfan-associated disorders. J. Hum. Genet. 52:1–12, 2007.

    Article  PubMed  CAS  Google Scholar 

  14. Peng, T., X. Lu, and Q. Feng. Pivotal role of gp91phox-containing NADH oxidase in lipopolysaccharide-induced tumor necrosis factor-alpha expression and myocardial depression. Circulation 111:1637–1644, 2005.

    Article  PubMed  CAS  Google Scholar 

  15. Person, A. D., S. E. Klewer, and R. B. Runyan. Cell biology of cardiac cushion development. Int. Rev. Cytol. 243:287–335, 2005.

    Article  PubMed  CAS  Google Scholar 

  16. Rabkin-Aikawa, E., J. E. Mayer, Jr., and F. J. Schoen. Heart valve regeneration. Adv. Biochem. Eng. Biotechnol. 94:141–179, 2005.

    PubMed  Google Scholar 

  17. Rabkin, E., and F. J. Schoen. Cardiovascular tissue engineering. Cardiovasc. Pathol. 11:305–317, 2002.

    Article  PubMed  Google Scholar 

  18. Vesely, I. Heart valve tissue engineering. Circ. Res. 97:743–755, 2005.

    Article  PubMed  CAS  Google Scholar 

  19. Warnock, J. N., et al. Design of a sterile organ culture system for the ex vivo study of aortic heart valves. J. Biomech. Eng. 127:857–861, 2005.

    Article  PubMed  Google Scholar 

  20. Wiechert, S., et al. 24-h Langendorff-perfused neonatal rat heart used to study the impact of adenoviral gene transfer. Am. J. Physiol. Heart Circ. Physiol. 285:H907–H914, 2003.

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank L. Emile for expert technical assistance. This study was supported by the American Heart Association (0555840T to V.G. and 0625861T to B.P.T.K.), the March of Dimes Birth Defects Foundation (1-FY06-375 to V.G.), and the NIH (5R21HL084278-02 to V.G.), NIH grants AG027211; HL033107; HL059139; HL069752; HL093481 (S.F.V) and the New Jersey Commission on Science and Technology through the New Jersey Center for Micro-Flow Control (N.A). Dr. Aubry is currently a Professor in the Department of Mechanical Engineering at Carnegie Mellon University, Pittsburgh, PA, USA.

Author information

Authors and Affiliations

  1. Department of Cell Biology and Molecular Medicine, Cardiovascular Research Institute, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, 185 South Orange Avenue, MSB Room G-609, Newark, NJ, 07103, USA

    Samuel C. Lieber, Boudewijn P. T. Kruithof, Stephen F. Vatner & Vinciane Gaussin

  2. Department of Mechanical Engineering, New Jersey Institute of Technology, Newark, NJ, USA

    Samuel C. Lieber & Nadine Aubry

Authors
  1. Samuel C. Lieber
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Boudewijn P. T. Kruithof
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nadine Aubry
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stephen F. Vatner
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vinciane Gaussin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stephen F. Vatner.

Additional information

Associate Editor Michael B. Lawrence oversaw the review of this article.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lieber, S.C., Kruithof, B.P.T., Aubry, N. et al. Design of a Miniature Tissue Culture System to Culture Mouse Heart Valves. Ann Biomed Eng 38, 674–682 (2010). https://doi.org/10.1007/s10439-010-9922-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10439-010-9922-8

Keywords

Search

Navigation