Three-dimensional engineered heart tissue from neonatal rat cardiac myocytes

Zimmermann, Wolfram H.; Fink, Christine; Kralisch, Dirk; Remmers, Ute; Weil, Joachim; Eschenhagen, Thomas

doi:10.1002/(sici)1097-0290(20000405)68:1<106::aid-bit13>3.0.co;2-3

Cited by 434 publications

(356 citation statements)

References 15 publications

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“…Depending on Re and the velocity entrance length, the flow within the channels will be either fully developed or plug flow. In our case, the flow was fully developed and the following equation was used to describe the profile: (12) U c is the average velocity through each channel determined as: (13) where N is the number of channels in the scaffold.…”

Section: Channeled Scaffolds With Pfc Oxygen Carriersmentioning

confidence: 99%

“…Measurement of contractile force is an additional useful functional assay 12 . We provide a detailed description of methods that we developed or that required significant modification compared to the standard protocol.…”

Section: Assessmentsmentioning

confidence: 99%

“…Inallof these systems, oxygen dissolved in culture medium was transported to the cells by molecular diffusion. Whereas human cardiac muscle is ~1 cm thick, diffusion alone can support only four to seven cell layers, that is, a 100-μm thick layer of viable and compact tissue 3,4,7,12 . We recently measured oxygen gradients in statically grown cardiac constructs and correlated them with the decrease in cell viability and density 13 (Fig.…”

Section: Introductionmentioning

confidence: 99%

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Cardiac tissue engineering using perfusion bioreactor systems

et al. 2008

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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 highfidelity models for biological research.

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Section: Channeled Scaffolds With Pfc Oxygen Carriersmentioning

confidence: 99%

Section: Assessmentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cardiac tissue engineering using perfusion bioreactor systems

et al. 2008

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“…The reconstitution of sufficiently strong contracting heart muscle constructs (considered to be the capacity to generate a systolic force of 1-3 mN/ mm 2 (Zimmermann et al, 2002) has proven difficult using solid scaffolds (Li et al, 1999). Liquid matrices provided improved functional properties of the constructs (Zimmermann et al, 2000). Scaffold-free cell sheet stacking after cultivation of monolayer patches on a thermosensitive culturing membrane showed promising mechanical properties (Shimizu et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…Numerous different tissue-like culture models are established by using various culture techniques, biological and nonbiological scaffolds, culture media, and cell sources (Zimmermann et al, 2000;Shimizu et al, 2003;Shin et al, 2003;Kelm et al, 2004;Kofidis et al, 2004). The aim of our study was to develop an easy-to-produce threedimensional scaffold-free culture model with homogenously distributed cardiomyocytes that could serve as a robust test system for basic research in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

Formation of three‐dimensional fetal myocardial tissue cultures from rat for long‐term cultivation

Just

Kürsten

Borth-Bruhns

et al. 2006

Developmental Dynamics

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Three-dimensional cardiomyocyte cultures offer new possibilities for the analysis of cardiac cell differentiation, spatial cellular arrangement, and time-specific gene expression in a tissue-like environment. We present a new method for generating homogenous and robust cardiomyocyte tissue cultures with good long-term viability. Ventricular heart cells prepared from fetal rats at embryonic day 13 were cultured in a scaffold-free two-step process. To optimize the cell culture model, several digestion protocols and culture conditions were tested. After digestion of fetal cardiac ventricles, the resultant cell suspension of isolated cardiocytes was shaken to initialize cell aggregate formation. In the second step, these three-dimensional cell aggregates were transferred onto a microporous membrane to allow further microstructure formation. Autonomously beating cultures possessed more than 25 cell layers and a homogenous distribution of cardiomyocytes without central necrosis after 8 weeks in vitro. The cardiomyocytes showed contractile elements, desmosomes, and gap junctions analyzed by immunohistochemistry and electron microscopy. The beat frequency could be modulated by adrenergic agonist and antagonist. Adenoviral green fluorescent protein transfer into cardiomyocytes was possible and highly effective. This three-dimensional tissue model proved to be useful for studying cell-cell interactions and cell differentiation processes in a three-dimensional cell arrangement. Developmental Dynamics 235: 2200 -2209, 2006.

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Cardiac Differentiation of Human Embryonic Stem Cells and their Assembly into Engineered Heart Muscle

2012

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The advent of pluripotent human embryonic stem cells has created the unique opportunity for the development of a wide variety of humanized cellular tools for basic research, as well as industrial and clinical applications. It has, however, become apparent that embryonic stem cell derivatives in classical monolayer or embryoid body culture do not resemble bona fide tissues, mainly because of their limited organotypic organization and maturation in these culture formats. This shortcoming may be addressed by tissue engineering technologies aiming at the provision of a “natural” growth environment to facilitate organotypic tissue assembly. In this unit, we provide two harmonized basic protocols for (1) cardiac differentiation of human embryonic stem cells under serum‐free conditions and (2) the assembly of the stem cell–derived cardiomyocytes into engineered heart muscle. This protocol can be easily adapted to bioengineer heart muscle also from other stem cell–derived cardiomyocytes, including cardiomyocytes from human‐induced pluripotent stem cells. Curr. Protoc. Cell Biol. 55:23.8.1‐23.8.21. © 2012 by John Wiley & Sons, Inc.

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Three-dimensional engineered heart tissue from neonatal rat cardiac myocytes

Cited by 434 publications

References 15 publications

Cardiac tissue engineering using perfusion bioreactor systems

Cardiac tissue engineering using perfusion bioreactor systems

Formation of three‐dimensional fetal myocardial tissue cultures from rat for long‐term cultivation

Cardiac Differentiation of Human Embryonic Stem Cells and their Assembly into Engineered Heart Muscle

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