Cardiac tissue engineering using perfusion bioreactor systems

This protocol describes tissue engineering of synchronously contractile cardiac constructs by culturing cardiac cell populations on porous scaffolds (in some cases with an array of channels) and bioreactors with perfusion of culture medium (in some cases supplemented with an oxygen carrier). The overall approach is 'biomimetic' in nature as it tends to provide in vivo-like oxygen supply to cultured cells and thereby overcome inherent limitations of diffusional transport in conventional culture systems. In order to mimic the capillary network, cells are cultured on channeled elastomer scaffolds that are perfused with culture medium that can contain oxygen carriers. The overall protocol takes 2–4 weeks, including assembly of the perfusion systems, preparation of scaffolds, cell seeding and cultivation, and on-line and end-point assessment methods. This model is well suited for a wide range of cardiac tissue engineering applications, including the use of human stem cells, and high-fidelity models for biological research.

[1]  Robert L Sah,et al.  Perfusion increases cell content and matrix synthesis in chondrocyte three-dimensional cultures. , 2002, Tissue engineering.

[2]  Milica Radisic,et al.  Oxygen gradients correlate with cell density and cell viability in engineered cardiac tissue , 2006, Biotechnology and bioengineering.

[3]  R Langer,et al.  Tissue engineering of functional cardiac muscle: molecular, structural, and electrophysiological studies. , 2001, American journal of physiology. Heart and circulatory physiology.

[4]  N. Stathopoulos,et al.  Shear stress effects on human embryonic kidney cells in Vitro , 1985, Biotechnology and bioengineering.

[5]  Gordana Vunjak-Novakovic,et al.  Effects of oxygen on engineered cardiac muscle. , 2002, Biotechnology and bioengineering.

[6]  Antonios G. Mikos,et al.  Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Antonios G. Mikos,et al.  Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Weisel,et al.  Construction of a bioengineered cardiac graft. , 2000, The Journal of thoracic and cardiovascular surgery.

[9]  P. Greenfield,et al.  Shear sensitivity of three hybridoma cell lines in suspension culture , 1987 .

[10]  R. Weisel,et al.  Survival and function of bioengineered cardiac grafts. , 1999, Circulation.

[11]  R J Cohen,et al.  Cardiac muscle tissue engineering : toward an in vitro model for electrophysiological studies , 1999 .

[12]  R. Nuccitelli.  Endogenous ionic currents and DC electric fields in multicellular animal tissues. , 1992, Bioelectromagnetics.

[13]  Tal Dvir,et al.  Activation of the ERK1/2 cascade via pulsatile interstitial fluid flow promotes cardiac tissue assembly. , 2007, Tissue engineering.

[14]  J. Kirkpatrick,et al.  Echocardiography in heart failure: applications, utility, and new horizons. , 2007, Journal of the American College of Cardiology.

[15]  Francesco Migliavacca,et al.  The effect of media perfusion on three-dimensional cultures of human chondrocytes: integration of experimental and computational approaches. , 2004, Biorheology.

[16]  Jin Gao,et al.  Macroporous elastomeric scaffolds with extensive micropores for soft tissue engineering. , 2006, Tissue engineering.

[17]  M. Radisic,et al.  Cell seeding of polymer scaffolds. , 2004, Methods in molecular biology.

[18]  Fen Chen,et al.  Biomimetic approach to cardiac tissue engineering: oxygen carriers and channeled scaffolds. , 2006, Tissue engineering.

[19]  Silviu Itescu,et al.  Cardiac Tissue Engineering in an In Vivo Vascularized Chamber , 2007, Circulation.

[20]  Gordana Vunjak-Novakovic,et al.  Perfusion improves tissue architecture of engineered cardiac muscle. , 2002, Tissue engineering.

[21]  R. Langer,et al.  A tough biodegradable elastomer , 2002, Nature Biotechnology.

[22]  Milica Radisic,et al.  Mathematical model of oxygen distribution in engineered cardiac tissue with parallel channel array perfused with culture medium containing oxygen carriers. , 2005, American journal of physiology. Heart and circulatory physiology.

[23]  F J Schoen,et al.  Cardiac tissue engineering: cell seeding, cultivation parameters, and tissue construct characterization. , 1999, Biotechnology and bioengineering.

[24]  R Langer,et al.  Biomimetic approach to cardiac tissue engineering , 2007, Philosophical Transactions of the Royal Society B: Biological Sciences.

[25]  K. Schügerl,et al.  Response of mammalian cells to shear stress , 1991, Applied Microbiology and Biotechnology.

[26]  Milica Radisic,et al.  Microfabricated poly(ethylene glycol) templates enable rapid screening of triculture conditions for cardiac tissue engineering. , 2009, Journal of biomedical materials research. Part A.

[27]  S R Gonda,et al.  Cardiac organogenesis in vitro: reestablishment of three-dimensional tissue architecture by dissociated neonatal rat ventricular cells. , 1999, Tissue engineering.

[28]  Milica Radisic,et al.  High-density seeding of myocyte cells for cardiac tissue engineering. , 2003, Biotechnology and bioengineering.

[29]  Robert E Guldberg,et al.  Effects of medium perfusion rate on cell-seeded three-dimensional bone constructs in vitro. , 2003, Tissue engineering.

[30]  D. Wendt,et al.  Oscillating perfusion of cell suspensions through three‐dimensional scaffolds enhances cell seeding efficiency and uniformity , 2003, Biotechnology and bioengineering.

[31]  J. Leor,et al.  Bioengineered Cardiac Grafts: A New Approach to Repair the Infarcted Myocardium? , 2000, Circulation.

[32]  Milica Radisic,et al.  Medium perfusion enables engineering of compact and contractile cardiac tissue. , 2004, American journal of physiology. Heart and circulatory physiology.

[33]  L. Bonassar,et al.  Comparison of Chondrogensis in Static and Perfused Bioreactor Culture , 2000, Biotechnology progress.

[34]  W. Zimmermann,et al.  Three-dimensional engineered heart tissue from neonatal rat cardiac myocytes. , 2000, Biotechnology and bioengineering.