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Assembly of a functional 3D primary cardiac construct using magnetic levitation

  • Received: 09 June 2016 Accepted: 15 July 2016 Published: 19 July 2016
  • Easily assembled organotypic co-cultures have long been sought in medical research. In vitro tissue constructs with faithful representation of in vivo tissue characteristics are highly desirable for screening and characteristic assessment of a variety of tissue types. Cardiac tissue analogs are particularly sought after due to the phenotypic degradation and difficulty of culture of primary cardiac myocytes. This study utilized magnetic nanoparticles and primary cardiac myocytes in order to levitate and culture multicellular cardiac aggregates (MCAs). Cells were isolated from 2 day old Sprague Dawley rat hearts and subsequently two groups were incubated with either C1: 33 µL nanoshell/million cells or C2: 50 µL nanoshell/million cells. Varying numbers of cells for each concentration were cultured in a magnetic field in a 24 well plate and observed over a period of 12 days. Constructs generally formed spherical structures. Masson’s trichrome staining of a construct shows the presence of extracellular matrix protein, indicating the presence of functional fibroblasts. Many constructs exhibited noticeable contraction after 4 days of culture and continued contracting noticeably past day 9 of culture. Noticeable contractility indicates the presence of functional primary cardiac myocytes in culture. Phenotypic conservation of cardiac cells was ascertained using IHC staining by α-actinin and collagen. CD31 and fibrinogen were probed in order to assess localization of fibroblasts and endothelial cells. The study verifies a protocol for the use of magnetic levitation in order to rapidly assemble 3D cardiac like tissue with phenotypic and functional stability.

    Citation: Matthew Hogan, Glauco Souza, Ravi Birla. Assembly of a functional 3D primary cardiac construct using magnetic levitation[J]. AIMS Bioengineering, 2016, 3(3): 277-288. doi: 10.3934/bioeng.2016.3.277

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  • Easily assembled organotypic co-cultures have long been sought in medical research. In vitro tissue constructs with faithful representation of in vivo tissue characteristics are highly desirable for screening and characteristic assessment of a variety of tissue types. Cardiac tissue analogs are particularly sought after due to the phenotypic degradation and difficulty of culture of primary cardiac myocytes. This study utilized magnetic nanoparticles and primary cardiac myocytes in order to levitate and culture multicellular cardiac aggregates (MCAs). Cells were isolated from 2 day old Sprague Dawley rat hearts and subsequently two groups were incubated with either C1: 33 µL nanoshell/million cells or C2: 50 µL nanoshell/million cells. Varying numbers of cells for each concentration were cultured in a magnetic field in a 24 well plate and observed over a period of 12 days. Constructs generally formed spherical structures. Masson’s trichrome staining of a construct shows the presence of extracellular matrix protein, indicating the presence of functional fibroblasts. Many constructs exhibited noticeable contraction after 4 days of culture and continued contracting noticeably past day 9 of culture. Noticeable contractility indicates the presence of functional primary cardiac myocytes in culture. Phenotypic conservation of cardiac cells was ascertained using IHC staining by α-actinin and collagen. CD31 and fibrinogen were probed in order to assess localization of fibroblasts and endothelial cells. The study verifies a protocol for the use of magnetic levitation in order to rapidly assemble 3D cardiac like tissue with phenotypic and functional stability.


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    [1] Ehler E, Jayasinghe SN (2014) Cell electrospinning cardiac patches for tissue engineering the heart. Analyst 139: 4449–52. doi: 10.1039/C4AN00766B
    [2] Haraguchi Y, Shimizu T, Matsuura K, et al. (2014) Cell sheet technology for cardiac tissue engineering. Methods Mol Biol 1181: 139c55.
    [3] Hirt MN, Hansen A, Eschenhagen T (2014) Cardiac tissue engineering: state of the art. Circ Res 114: 354–67.
    [4] Macadangdang J, Lee HJ, Carson D, et al. (2014) Capillary force lithography for cardiac tissue engineering. J Vis Exp (88): 50039.
    [5] Matsuura K, Masuda S, Shimizu T (2014) Cell sheet-based cardiac tissue engineering. AnatRec(Hoboken) 297: 65–72.
    [6] Rao C, Barratt H, Prodromakis T, et al. (2014) Tissue engineering techniques in cardiac repair and disease modelling. Curr Pharm Des 20: 2048–56. doi: 10.2174/13816128113199990442
    [7] Sapir Y, Polyak B, Cohen S (2014) Cardiac tissue engineering in magnetically actuated scaffolds. Nanotechnology 25: 014009. doi: 10.1088/0957-4484/25/1/014009
    [8] Williams C, Budina E, Stoppel WL, et al. (2014) Cardiac Extracellular Matrix-Fibrin Hybrid Scaffolds with Tunable Properties for Cardiovascular Tissue Engineering. Acta Biomater 14: 84–95.
    [9] Galvez-Monton C, Prat-Vidal C, Roura S, et al. (2013) Cardiac Tissue Engineering and the Bioartificial Heart. Rev Esp Cardiol 66: 391–9. doi: 10.1016/j.recesp.2012.11.013
    [10] Roger VL, Go AS, Lloyd-Jones DM, et al. (2012) Heart disease and stroke statistics--2012 update: a report from the American Heart Association. Circulation 125: e2–e220. doi: 10.1161/CIR.0b013e31823ac046
    [11] Tao ZW, Mohamed M, Hogan M, et al. (2014) Optimizing a spontaneously contracting heart tissue patch with rat neonatal cardiac cells on fibrin gel. J Tissue Eng Regen Med. doi: 10.1002/term.1895.
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  • © 2016 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)
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