Development of three-dimensional primary human myospheres as culture model of skeletal muscle cells for metabolic studies

Andrea Dalmao-Fernandez; Aleksandra Aizenshtadt; Hege G Bakke; Stefan Krauss; Arild C Rustan; G Hege Thoresen; Eili Tranheim Kase

doi:10.3389/fbioe.2023.1130693

Development of three-dimensional primary human myospheres as culture model of skeletal muscle cells for metabolic studies

Front Bioeng Biotechnol. 2023 Mar 23:11:1130693. doi: 10.3389/fbioe.2023.1130693. eCollection 2023.

Authors

Andrea Dalmao-Fernandez¹, Aleksandra Aizenshtadt², Hege G Bakke¹, Stefan Krauss², Arild C Rustan¹, G Hege Thoresen^{1

3}, Eili Tranheim Kase¹

Affiliations

¹ Section for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, University of Oslo, Oslo, Norway.
² Hybrid Technology Hub Centre of Excellence, Faculty of Medicine, University of Oslo, Oslo, Norway.
³ Department of Pharmacology, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.

Abstract

Introduction: Skeletal muscle is a major contributor to whole-body energy homeostasis and the utilization of fatty acids and glucose. At present, 2D cell models have been the most used cellular models to study skeletal muscle energy metabolism. However, the transferability of the results to in vivo might be limited. This project aimed to develop and characterize a skeletal muscle 3D cell model (myospheres) as an easy and low-cost tool to study molecular mechanisms of energy metabolism. Methods and results: We demonstrated that human primary myoblasts form myospheres without external matrix support and carry structural and molecular characteristics of mature skeletal muscle after 10 days of differentiation. We found significant metabolic differences between the 2D myotubes model and myospheres. In particular, myospheres showed increased lipid oxidative metabolism than the 2D myotubes model, which oxidized relatively more glucose and accumulated more oleic acid. Discussion and conclusion: These analyses demonstrate model differences that can have an impact and should be taken into consideration for studying energy metabolism and metabolic disorders in skeletal muscle.

Keywords: 3D cell model; energy metabolism; metabolic disorders; muscle spheroid; myosphere; skeletal muscle.

Grants and funding

This work received funding from the European Union’s Horizon 2020 Research and Innovation program under the Marie Skłodowska-Curie Actions Grant, agreement No. 80113 (Scientia fellowship), and from the Research Council of Norway, through its Centers of Excellence scheme; project No. 262613.