In vitro-generated human muscle reserve cells are heterogeneous for Pax7 with distinct molecular states and metabolic profiles

Article Article scientifique Article In vitro-generated human muscle reserve cells are heterogeneous for Pax7 with distinct molecular states and metabolic profiles eng 2023-09-08 2023-09-08 author Bouche Axelle Annie 0009-0008-6188-2509 Cell Therapy and Musculoskeletal Disorders Laboratory, Department of Orthopedic Surgery, Geneva University Hospitals and Faculty of Medicine, University Medical Center, 1 rue Michel Servet, 1211, Geneva, Switzerland.,Department of Cell Physiology and Metabolism, Faculty of Medicine, Geneva, Switzerland. 931839 Département de chirurgie (CHIRU) author Borner Benoît Cell Therapy and Musculoskeletal Disorders Laboratory, Department of Orthopedic Surgery, Geneva University Hospitals and Faculty of Medicine, University Medical Center, 1 rue Michel Servet, 1211, Geneva, Switzerland. 677342 author Richard Chloé Cell Therapy and Musculoskeletal Disorders Laboratory, Department of Orthopedic Surgery, Geneva University Hospitals and Faculty of Medicine, University Medical Center, 1 rue Michel Servet, 1211, Geneva, Switzerland.,Department of Cell Physiology and Metabolism, Faculty of Medicine, Geneva, Switzerland. 899843 author Grand Ysaline Cell Therapy and Musculoskeletal Disorders Laboratory, Department of Orthopedic Surgery, Geneva University Hospitals and Faculty of Medicine, University Medical Center, 1 rue Michel Servet, 1211, Geneva, Switzerland.,Department of Cell Physiology and Metabolism, Faculty of Medicine, Geneva, Switzerland. 834072 author Hannouche Didier 0000-0003-2060-2394 Cell Therapy and Musculoskeletal Disorders Laboratory, Department of Orthopedic Surgery, Geneva University Hospitals and Faculty of Medicine, University Medical Center, 1 rue Michel Servet, 1211, Geneva, Switzerland.,Department of Cell Physiology and Metabolism, Faculty of Medicine, Geneva, Switzerland. 842478 Département de chirurgie (CHIRU) author Laumonier Thomas 0000-0002-8662-2972 Cell Therapy and Musculoskeletal Disorders Laboratory, Department of Orthopedic Surgery, Geneva University Hospitals and Faculty of Medicine, University Medical Center, 1 rue Michel Servet, 1211, Geneva, Switzerland. thomas.laumonier@unige.ch.,Department of Cell Physiology and Metabolism, Faculty of Medicine, Geneva, Switzerland. thomas.laumonier@unige.ch. 135589 Département de chirurgie (CHIRU) 243 10.1186/s13287-023-03483-5 37679820 1757-6512 PMC10486062 Background: The capacity of skeletal muscles to regenerate relies on Pax7+muscle stem cells (MuSC). While in vitro-amplified MuSC are activated and lose part of their regenerative capacity, in vitro-generated human muscle reserve cells (MuRC) are very similar to quiescent MuSC with properties required for their use in cell-based therapies. Methods: In the present study, we investigated the heterogeneity of human MuRC and characterized their molecular signature and metabolic profile. Results: We observed that Notch signaling is active and essential for the generation of quiescent human Pax7+MuRC in vitro. We also revealed, by immunofluorescence and flow cytometry, two distinct subpopulations of MuRC distinguished by their relative Pax7 expression. After 48 h in differentiation medium (DM), the Pax7Highsubpopulation represented 35% of the total MuRC pool and this percentage increased to 61% after 96 h in DM. Transcriptomic analysis revealed that Pax7HighMuRC were less primed for myogenic differentiation as compared to Pax7LowMuRC and displayed a metabolic shift from glycolysis toward fatty acid oxidation. The bioenergetic profile of human MuRC displayed a 1.5-fold decrease in glycolysis, basal respiration and ATP-linked respiration as compared to myoblasts. We also observed that AMPKα1 expression was significantly upregulated in human MuRC that correlated with an increased phosphorylation of acetyl-CoA carboxylase (ACC). Finally, we showed that fatty acid uptake was increased in MuRC as compared to myoblasts, whereas no changes were observed for glucose uptake. Conclusions: Overall, these data reveal that the quiescent MuRC pool is heterogeneous for Pax7 with a Pax7Highsubpopulation being in a deeper quiescent state, less committed to differentiation and displaying a reduced metabolic activity. Altogether, our data suggest that human Pax7HighMuRC may constitute an appropriate stem cell source for potential therapeutic applications in skeletal muscle diseases. AMPK Fatty acid oxidation Human muscle reserve cell Metabolism Pax7 heterogeneity Quiescence Humans Muscle Cells Satellite Cells, Skeletal Muscle Muscle, Skeletal Fatty Acids Metabolome Dewey 612 Dewey 617 57 Département de chirurgie 222 134 Département de physiologie cellulaire et métabolisme 21C false 955 Mécanismes physiopathologiques de l'arthrose 960 https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE233597 Stem cell research & therapy 14 1 ec24830b-5dfe-401d-b6d6-34b60ac41c57 Article (Published version) Public Article-scientifique-from-europePmc.pdf 2672881 application/pdf CC BY-4.0 eb65b895d649588bce9742eb123dcdbf48d950a4e0a30157d94e6fa99b86173e e117af29-ef1b-4523-a9f6-2e089097785f Appendix Public 13287_2023_3483_MOESM1_ESM.docx 747267 application/vnd.openxmlformats-officedocument.wordprocessingml.document ae6f18542adea5784428b4d7e67b0317913e05f6c6d96efe51e5224d248565ea openAccess Faculté de médecine / Section de médecine clinique / Département de chirurgie Faculté de médecine / Section de médecine fondamentale / Département de physiologie cellulaire et métabolisme

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In vitro-generated human muscle reserve cells are heterogeneous for Pax7 with distinct molecular states and metabolic profiles	Stem cell research & therapy			2023	206	35

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