Electrophysiologically competent skeletal myoblast based engineered tissue constructs show cardiac-like force generation

Klaus Neef, A.-C. Deppe, Sureshkumar Perumal Srinivasan, Philipp Treskes, Christof Stamm, Thorsten Wahlers, Yeong-Hoon Choi

Research output: Contribution to conferenceAbstractpeer-review

Abstract

Introduction: Previously, we demonstrated that skeletal myoblast (SM) based engineered tissue constructs (ETC) applying specific culture conditions can augment the intra-cardiac conduction system, i.e. for the treament of AV conduction block. However, the mechanical and contractile properties of SM based ETC remain largely unexplored. Here, we present force measurement analyses of ETC as an essential prerequisite for potential treatment of the failing heart.

Methods: SM were prepared from skeletal muscle tissue, expanded in cell culture and analyzed for cell type specific protein expression. For the fabrication of ETC, purified SM cell populations were seeded into hydrocollagen matrices. After casting the cell/matrix mixture in specific molds polymerization led to generation of passive longitudinal force within ETC. After submission to cell culture conditions which enabled the ETC to form myotubes, ETC were analyzed histologically. Contractile function was determined using a one-dimensional force transducer, with respect to stimulation frequency, maximum force generation, tetanic force generation and muscular fatiguing. Neonatal skeletal muscle and myocardium were used as references to evaluate specificity of force generation.

Results: ETC containing 5x10E6 cell cultured expanded SM with at a purity of 92.3 ± 5.4% desmin positive cells, were submitted to cell culture conditions resulting in longitudinal organization of single cells and could be successfully differentiated to syncytic myotubes. Force measurement revealed an active contractile force of 1143 ± 289 µN for ETC (length: 0.9 cm; diameter: 1 mm). There was no significant change in maximum force generation for stimulation frequencies from 0.2 to 10 Hz. Applying constant stimulation at 15 Hz a decrease in maximum force (-16.7 ± 6.1%) was observed after 1 minute. This was comparable to measurements of neonatal cardiac muscle (max force: 1975 ± 214 µN; constant force from 0.2-40 Hz; all p>0.05), but significantly different from measurements of neonatal skeletal muscle (max force: 3953 ± 871 µN; constant force from 0.2-5 Hz; decrease in maximum force: -53.7 ± 26.2% after 1 minute; all p < 0.05).

Conclusion: SM based ETC exhibit force generating properties similar to myocardial tissue. In addition to the previous electrophysiological results, these data re-confirm SM as potential substrate for cardiac cell therapy.
Original languageEnglish
Publication statusPublished - Oct 2014
Event46th Annual Meeting of the German Society for Pediatric Cardiology (DGPK) - Weimar, Germany
Duration: 4 Oct 20147 Oct 2014

Conference

Conference46th Annual Meeting of the German Society for Pediatric Cardiology (DGPK)
Country/TerritoryGermany
CityWeimar
Period4/10/147/10/14

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