Dataset Information

The use of three-dimensional primary human myospheres to explore skeletal muscle effects of in vivo krill oil supplementation

ABSTRACT: Supplementation with krill oil has shown effects on whole-body lipid and glucose metabolism, as well as on skeletal muscle strength and function. The most widely used model for culturing skeletal muscle cells (myotubes) is two-dimensional (2D), however there is growing interest in establishing 3D muscle cell models. We previously showed that intervention with krill oil in vivo promoted increased fatty acid metabolism and protein synthesis in cultured human myotubes in a 2D model. The aim of this study was to explore the effects of krill oil supplementation in vivo in a 3D myosphere model to evaluate the usefulness of a human skeletal 3D model compared to established 2D models. Myospheres were formed from myoblasts obtained before and after 7 weeks of in vivo krill oil intervention. Glucose and oleic acid metabolism were assessed, and transcriptomic and proteomic analyses were also performed. In contrast to previous data found in 2D myotube cultures, in vivo intervention with krill oil increased glucose metabolism in myospheres, while no effect was observed on fatty acid metabolism. Transcriptomic and proteomic analyses in myospheres obtained before and after krill oil supplementation showed differentially expressed genes/proteins involved in pathways like motor proteins, hypertrophy, calcium signaling and glycolysis/gluconeogenesis, of which motor proteins and hypertrophy have not been described in 2D myotube cultures. These findings suggested that in vivo krill oil intervention induces a different metabolic effect when comparing 3D and 2D cultures. In addition, in contrast to the 2D model, data obtained with the 3D model showed gene expression changes that are more compatible with previously observed results in vivo concerning skeletal muscle motoric function. Hence, the 3D model might better reflect krill oil-induced modifications in skeletal muscle performance in vivo than the 2D model.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Tuula Nyman

LAB HEAD: Tuula Nyman

PROVIDER: PXD055840 | Pride | 2025-08-04

REPOSITORIES: Pride

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The use of three-dimensional primary human myospheres to explore skeletal muscle effects of in vivo krill oil supplementation.

Dalmao-Fernandez Andrea A Katare Parmeshwar B PB Bakke Hege G HG Hamarsland Håvard H Ellefsen Stian S Singh Sachin S Nyman Tuula Anneli TA Kase Eili Tranheim ET Rustan Arild C AC Thoresen G Hege GH

In vitro models 20250430 2

<h4>Purpose</h4>Supplementation with krill oil has shown effects on whole-body lipid and glucose metabolism, as well as on skeletal muscle strength and function. We previously showed that krill oil intervention in vivo promoted fatty acid metabolism and protein synthesis in cultured human myotubes in a two-dimensional (2D) model. The aim of this study was to explore the effects of krill oil supplementation in vivo in a 3D myosphere model, and to compare a the human skeletal muscle 3D cell model ...[more]

PMID: 40708816

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Project description:Purpose: Supplementation with krill oil has shown effects on whole-body lipid and glucose metabolism, as well as on skeletal muscle strength and function. We previously showed that krill oil intervention in vivo promoted fatty acid metabolism and protein synthesis in cultured human myotubes in a two-dimensional (2D) model. The aim of this study was to explore the effects of krill oil supplementation in vivo in a 3D myosphere model and to compare the 3D and 2D models. Methods: Myospheres were formed from myoblasts obtained before and after 7 weeks of in vivo krill oil intervention. Glucose and oleic acid metabolism were assessed, and transcriptomic and proteomic analyses were performed. Results: In vivo intervention with krill oil increased glucose metabolism in myospheres, while no effect was observed on fatty acid metabolism. Transcriptomic analyses of myospheres after krill oil intervention showed increased expression of genes involved in pathways like motor proteins and hypertrophy, as well as in calcium signaling, of which motor proteins and hypertrophy pathways have not been described in 2D myotube cultures. Proteomic analyses after krill oil intervention showed increased expression of proteins in glycolysis/gluconeogenesis and fatty acid degradation. Comparison of proteins expressed in the 3D myosphere model and a 2D myotube model at the basal level showed that in myospheres, mitochondrial gene expression and translation dominated, while in 2D cultures, mitochondrial organization and response to oxidative stress were more important. Conclusion: These findings suggest that in vivo krill oil intervention induces different metabolic effects when comparing 3D and 2D cultures. In contrast to the 2D model, data obtained with the 3D model showed gene expression changes that are more compatible with previously observed results in vivo concerning skeletal muscle motoric function. Hence, the 3D model might better reflect krill oil-induced modifications in skeletal muscle performance in vivo than the 2D model.

Project description:Background Three-dimensional (3D) bioengineered models of human skeletal muscle are a promising approach for studying muscle development, function and disease in vitro. These models more closely resemble the complexity of native muscle tissue than two-dimensional (2D) monolayer culture and allow for functional measurements to be performed. However, a more complete understanding of how culture condition and duration impacts the myotube maturity and function is required to validate the transition from 2D to 3D culture of muscle cells. Methods Human skeletal muscle cells were cultured as either 2D monolayers or within 3D fibrin-based hydrogels as muscle constructs for up to 21 days. Quantitative proteomic analysis and functional assessments, including contractile force and cross-sectional area measurements, were conducted to evaluate the impact of culture conditions and duration on muscle cell differentiation. Results Proteomic analysis revealed myoblasts differentiated into myotubes by 8 days of differentiation in both 2D and 3D environments. However, the proteomic profiles of myotubes varied significantly between the two culture environments. At day 8 of differentiation, myosin heavy chain isotype abundance indicated a predominantly slow-twitch phenotype in 3D constructs, compared to a mixed fibre type phenotype in 2D monolayers. By day 21 of differentiation, 3D muscle constructs displayed improved mitochondrial maturity, extracellular matrix remodelling, and signs of transitioning towards a fast-twitch phenotype. This prolonged culture duration also resulted in increased passive tension but decreased peak contractile force in 3D muscle constructs. Conclusions This study demonstrates that 3D culture promotes maturity in human skeletal muscle cells, mimicking the biochemical cues and energy demands seen in native muscle tissue. The data highlights the importance of selecting appropriate culture conditions and durations when studying skeletal muscle cells in vitro and suggests 8 days of differentiation as optimal for achieving peak contractile force in 3D muscle constructs.