Microfluidic patterning for fabrication of contractile cardiac organoids

The development of in vitro methods of engineering three-dimensional cardiac tissues can be useful for tissue replacement, diagnostics and drug discovery. Here, we introduce the use of patterned hyaluronic acid (HA) substrates generated using microfluidic patterning as a method of fabricating 3D cardiac organoids. HA micropatterns served as inductive templates for organoid assembly. Upon seeding, cardiomyocytes elongated and aligned along the pattern direction attaching preferentially to the glass substrate and the interface between HA patterns and glass substrate. After 3 days in culture, the linearly aligned myocytes detached from the surface and formed contractile cardiac organoids. The procedure can be utilized to simply, rapidly and inexpensively create in vitro cardiac tissue models.

[1]  Todd C McDevitt,et al.  In vitro generation of differentiated cardiac myofibers on micropatterned laminin surfaces. , 2002, Journal of biomedical materials research.

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

[3]  G. Whitesides,et al.  Self-assembled organic monolayers: model systems for studying adsorption of proteins at surfaces , 1991, Science.

[4]  Daniel I. C. Wang,et al.  Engineering cell shape and function. , 1994, Science.

[5]  M L Yarmush,et al.  Controlling cell interactions by micropatterning in co-cultures: hepatocytes and 3T3 fibroblasts. , 1997, Journal of biomedical materials research.

[6]  Y. Shimabukuro,et al.  Regeneration of periodontal tissues by basic fibroblast growth factor. , 1999, Journal of periodontal research.

[7]  Tejal A Desai,et al.  Microfluidic patterning of cells in extracellular matrix biopolymers: effects of channel size, cell type, and matrix composition on pattern integrity. , 2003, Tissue engineering.

[8]  K Rakusan,et al.  The effect of growth and aging on functional capillary supply of the rat heart. , 1982, Growth.

[9]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

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

[11]  Gordana Vunjak-Novakovic,et al.  Bioreactors mediate the effectiveness of tissue engineering scaffolds , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[12]  Ravi Birla,et al.  Self‐organization of rat cardiac cells into contractile 3‐D cardiac tissue , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[13]  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.

[14]  M. Toner,et al.  Microfabrication of Hepatocyte/Fibroblast Co‐cultures: Role of Homotypic Cell Interactions , 1998, Biotechnology progress.

[15]  A. Chilkoti,et al.  Microstamping on an Activated Polymer Surface: Patterning Biotin and Streptavidin onto Common Polymeric Biomaterials , 2001 .

[16]  G. Whitesides,et al.  Patterning Mammalian Cells Using Elastomeric Membranes , 2000 .

[17]  G M Whitesides,et al.  Patterning cells and their environments using multiple laminar fluid flows in capillary networks. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[18]  W. Zimmermann,et al.  Tissue Engineering of a Differentiated Cardiac Muscle Construct , 2002, Circulation research.

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

[20]  G M Whitesides,et al.  Using self-assembled monolayers to understand the interactions of man-made surfaces with proteins and cells. , 1996, Annual review of biophysics and biomolecular structure.

[21]  J E Saffitz,et al.  Synthetic Strands of Neonatal Mouse Cardiac Myocytes: Structural and Electrophysiological Properties , 2000, Circulation research.

[22]  T. Desai,et al.  Inhibition of fibroblast proliferation in cardiac myocyte cultures by surface microtopography. , 2003, American journal of physiology. Cell physiology.

[23]  F. Lapicque,et al.  Hyaluronate-alginate gel as a novel biomaterial: mechanical properties and formation mechanism. , 1999, Biotechnology and bioengineering.

[24]  Won-Gun Koh,et al.  Molding of hydrogel microstructures to create multiphenotype cell microarrays. , 2003, Analytical chemistry.

[25]  Aliasger K Salem,et al.  Rapid localized cell trapping on biodegradable polymers using cell surface derivatization and microfluidic networking. , 2006, Biomaterials.

[26]  G. Whitesides,et al.  Patterned deposition of cells and proteins onto surfaces by using three-dimensional microfluidic systems. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M L Yarmush,et al.  Probing heterotypic cell interactions: hepatocyte function in microfabricated co-cultures. , 1998, Journal of biomaterials science. Polymer edition.

[28]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[29]  G. Abatangelo,et al.  New frontiers in medical sciences : redefining hyaluronan : proceedings of the symposium held in Padua, Italy, 17-19 June 1999 , 2000 .

[30]  Mitsuo Umezu,et al.  Fabrication of Pulsatile Cardiac Tissue Grafts Using a Novel 3-Dimensional Cell Sheet Manipulation Technique and Temperature-Responsive Cell Culture Surfaces , 2002, Circulation research.

[31]  Ralph G Nuzzo,et al.  Engineering the morphology and electrophysiological parameters of cultured neurons by microfluidic surface patterning , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[32]  T. Desai,et al.  Fabrication of microtextured membranes for cardiac myocyte attachment and orientation. , 2000, Journal of biomedical materials research.

[33]  M. Morra.  Engineering of biomaterials surfaces by hyaluronan. , 2005, Biomacromolecules.

[34]  D. Fingar,et al.  The mTOR/p70 S6K1 pathway regulates vascular smooth muscle cell differentiation. , 2004, American journal of physiology. Cell physiology.

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

[36]  A. Khademhosseini,et al.  Soft Lithographic Patterning of Hyaluronic Acid on Hydrophilic Substrates Using Molding and Printing , 2004 .

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

[38]  R E Ideker,et al.  Nonlinear changes of transmembrane potential caused by defibrillation shocks in strands of cultured myocytes. , 2000, American journal of physiology. Heart and circulatory physiology.

[39]  D E Ingber,et al.  Using microcontact printing to pattern the attachment of mammalian cells to self-assembled monolayers of alkanethiolates on transparent films of gold and silver. , 1997, Experimental cell research.

[40]  D. Piacquadio,et al.  Evaluation of hylan b gel as a soft-tissue augmentation implant material. , 1997, Journal of the American Academy of Dermatology.

[41]  M L Yarmush,et al.  Effect of cell–cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[42]  D J Beebe,et al.  Microfabricated elastomeric stencils for micropatterning cell cultures. , 2000, Journal of biomedical materials research.

[43]  Tejal A Desai,et al.  Layer-by-layer microfluidics for biomimetic three-dimensional structures. , 2004, Biomaterials.

[44]  Gregory T. A. Kovacs,et al.  A closed-loop electrical stimulation system for cardiac cell cultures , 2005, IEEE Transactions on Biomedical Engineering.

[45]  Gordana Vunjak-Novakovic,et al.  Cultivation in rotating bioreactors promotes maintenance of cardiac myocyte electrophysiology and molecular properties. , 2003, Tissue engineering.

[46]  A. Kleber,et al.  Patterned growth of neonatal rat heart cells in culture. Morphological and electrophysiological characterization. , 1991, Circulation research.

[47]  D. Aeschlimann,et al.  New strategy for chemical modification of hyaluronic acid: preparation of functionalized derivatives and their use in the formation of novel biocompatible hydrogels. , 1999, Journal of biomedical materials research.

[48]  Milica Radisic,et al.  Functional assembly of engineered myocardium by electrical stimulation of cardiac myocytes cultured on scaffolds , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[49]  M. Miragoli,et al.  Coupling of Cardiac Electrical Activity Over Extended Distances by Fibroblasts of Cardiac Origin , 2003, Circulation research.

[50]  Hongwei Ma,et al.  Universal Route to Cell Micropatterning Using an Amphiphilic Comb Polymer , 2003 .

[51]  Ali Khademhosseini,et al.  Layer-by-layer deposition of hyaluronic acid and poly-L-lysine for patterned cell co-cultures. , 2004, Biomaterials.

[52]  D E Ingber,et al.  Controlling cell attachment on contoured surfaces with self-assembled monolayers of alkanethiolates on gold. , 1996, Proceedings of the National Academy of Sciences of the United States of America.