Maintenance of highly contractile tissue-cultured avian skeletal myotubes in collagen gel

Vandenburgh, Herman H.; Karlisch, Patricia; Farr, Lynne

doi:10.1007/bf02623542

Cited by 173 publications

(135 citation statements)

References 28 publications

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“…A number of recent studies have found that passive tension can be attributed to the cells' acting on the gel to generate the passive forces (7,9). The primary human muscle cells used in the present work caused gel contraction and dehydration as previously observed with avian and rodent muscle cells (38,40). This suggests that HBAMs are under passive tension in their normal tissue-cultured state.…”

Section: Discussionsupporting

confidence: 73%

Mechanical stimulation improves tissue-engineered human skeletal muscle

Powell

Smiley

Mills

et al. 2002

American Journal of Physiology-Cell Physiology

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397

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Human bioartificial muscles (HBAMs) are tissue engineered by suspending muscle cells in collagen/MATRIGEL, casting in a silicone mold containing end attachment sites, and allowing the cells to differentiate for 8 to 16 days. The resulting HBAMs are representative of skeletal muscle in that they contain parallel arrays of postmitotic myofibers; however, they differ in many other morphological characteristics. To engineer improved HBAMs, i.e., more in vivo-like, we developed Mechanical Cell Stimulator (MCS) hardware to apply in vivo-like forces directly to the engineered tissue. A sensitive force transducer attached to the HBAM measured real-time, internally generated, as well as externally applied, forces. The muscle cells generated increasing internal forces during formation which were inhibitable with a cytoskeleton depolymerizer. Repetitive stretch/relaxation for 8 days increased the HBAM elasticity two- to threefold, mean myofiber diameter 12%, and myofiber area percent 40%. This system allows engineering of improved skeletal muscle analogs as well as a nondestructive method to determine passive force and viscoelastic properties of the resulting tissue.

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Section: Discussionsupporting

confidence: 73%

Mechanical stimulation improves tissue-engineered human skeletal muscle

Powell

Smiley

Mills

et al. 2002

American Journal of Physiology-Cell Physiology

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397

385

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“…Collagen gel, however, is not suitable as scaffold for maturating muscle cell cultures. 29,37 Therefore, alternative scaffolds based on the work of Vandenburgh et al 39 are being developed for future studies. 8 Although the muscle cell/agarose system is a limited representation of in vivo muscle tissue it represents a simple, reproducible model to test our hypothesis on the damaging effects of compressive strain-induced muscle cell deformation.…”

Section: Discussionmentioning

confidence: 99%

Compressive Deformation and Damage of Muscle Cell Subpopulations in a Model System

Bouten

Knight

Lee

et al. 2001

Annals of Biomedical Engineering

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Abstract-To study the effects of compressive straining on muscle cell deformation and damage an in vitro model system was developed. Myoblasts were seeded in agarose constructs and cultured in growth medium for 4 days. Subsequently, the cells were allowed to fuse into multinucleated myotubes for 8 days in differentiation medium, resulting in a population of spherical myoblasts ͑50%͒, spherical myotubes ͑35%͒, and elongated myotubes ͑15%͒ with an overall viability of 90%. To evaluate cell deformation upon construct compression half-core shaped constructs were compressed up to 40% strain and the resulting cell shape was assessed from confocal scans through the central plane of spherical cells. The ratio of cell diameters measured parallel and perpendicular to the axis of compression was used as an index of deformation ͑DI͒. The average DI of myoblasts decreased with strain level ͑0.99Ϯ0.03, 0.70Ϯ0.04, and 0.56Ϯ0.10 at 0%, 20%, and 40% strain͒, whereas for myotubes DI decreased up to 20% strain and then remained fairly constant ͑0.99Ϯ0.06, 0.55Ϯ0.06, 0.50Ϯ0.11͒. The discrepancy in DI between spherical myoblasts and myotubes at 20% strain was explained by the relative sensitivity of the cell membrane to buckling, which is more pronounced in the myotubes. Sustained compression up to 24 h at 20% strain resulted in a significant increase in cell damage with time as compared to unstrained controls. Despite differences in membrane buckling no difference in damage between myoblasts and spherical myotubes was observed over time, whereas the elongated myotubes were more susceptible to damage.

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“…The hydrogel method has been pioneered in the 1980s as an advanced culture method for fibroblasts and skeletal muscle cells 6 and gave rise to the first successful cardiac tissue.…”

Section: Hydrogel Techniquementioning

confidence: 99%

Cardiac Tissue Engineering

Hirt

Hansen

Eschenhagen

2014

Circulation Research

355

290

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Abstract:The engineering of 3-dimensional (3D) heart muscles has undergone exciting progress for the past decade.Profound advances in human stem cell biology and technology, tissue engineering and material sciences, as well as prevascularization and in vitro assay technologies make the first clinical application of engineered cardiac tissues a realistic option and predict that cardiac tissue engineering techniques will find widespread use in the preclinical research and drug development in the near future. Tasks that need to be solved for this purpose include standardization of human myocyte production protocols, establishment of simple methods for the in vitro vascularization of 3D constructs and better maturation of myocytes, and, finally, thorough definition of the predictive value of these methods for preclinical safety pharmacology. The present article gives an overview of the present state of the art, bottlenecks, and perspectives of cardiac tissue engineering for cardiac repair and in vitro testing. ( Jeffrey Robbins, EditorOriginal received May 24, 2013; revision received July 12, 2013; accepted July 15, 2013. In November 2013, the average time from submission to first decision for all original research papers submitted to Circulation Research was 14.6 days.From the Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; and DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Hamburg, Germany.Correspondence to Thomas Eschenhagen, Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Martinistraße 52, 20246 Hamburg, Germany. E-mail t.eschenhagen@uke.de by guest on May 9, 2018 http://circres.ahajournals.org/ Downloaded from Hirt et al Cardiac Tissue Engineering 355gyratory shaking of embryonic chicken cardiac myocytes in an Erlenmeyer flask induced the formation of spontaneously beating spheroids with improved functionality when compared with standard 2D-cultured myocytes. This selfassembly is still used in variations to generate cardiac microspheres. 5 Current cardiac tissue engineering methods embark on this endogenous capacity and use engineering techniques to make 3D constructs of the desired size, geometry, and orientation (Table). Hydrogel TechniqueThe hydrogel method has been pioneered in the 1980s as an advanced culture method for fibroblasts and skeletal muscle cells 6 and gave rise to the first successful cardiac tissue.7 It needs 3 factors: solutions of gelling natural products, such as collagen I, matrigel, fibrin, or mixtures of them; casting molds; and anchoring constructs. The hydrogel entraps cells in a 3D space during gelling, the mold gives the 3D form, and the anchors allow the growing tissue to fix and to develop mechanical tension between ≥2 anchor points. Important aspects of this technique are that the naturally occurring hydrogels stimulate cells to spread and form intercellular connections and that the cells compress the gel, reduce the w...

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Maintenance of highly contractile tissue-cultured avian skeletal myotubes in collagen gel

Cited by 173 publications

References 28 publications

Mechanical stimulation improves tissue-engineered human skeletal muscle

Mechanical stimulation improves tissue-engineered human skeletal muscle

Compressive Deformation and Damage of Muscle Cell Subpopulations in a Model System

Cardiac Tissue Engineering

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