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Isolation and characterization of muscle stem cells, fibro-adipogenic progenitors, and macrophages from human skeletal muscle biopsies.
Jensen, Jonas B; Møller, Andreas B; Just, Jesper; Mose, Maike; de Paoli, Frank V; Billeskov, Tine B; Fred, Rikard G; Pers, Tune H; Pedersen, Steen B; Petersen, Klaus K; Bjerre, Mette; Farup, Jean; Jessen, Niels.
Afiliación
  • Jensen JB; Department of Biomedicine, Aarhus University, Aarhus, Denmark.
  • Møller AB; Research Laboratory for Biochemical Pathology, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
  • Just J; Steno Diabetes Center Aarhus, Aarhus University Hospital, Aarhus, Denmark.
  • Mose M; Research Laboratory for Biochemical Pathology, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
  • de Paoli FV; Steno Diabetes Center Aarhus, Aarhus University Hospital, Aarhus, Denmark.
  • Billeskov TB; Center of Functionally Integrative Neuroscience, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
  • Fred RG; Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark.
  • Pers TH; Medical Research Laboratory, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
  • Pedersen SB; Diabetes and Hormonal Diseases, Aarhus University Hospital, Aarhus, Denmark.
  • Petersen KK; Department of Biomedicine, Aarhus University, Aarhus, Denmark.
  • Bjerre M; Department of Cardiothoracic and Vascular Surgery, Aarhus University Hospital, Aarhus, Denmark.
  • Farup J; Research Laboratory for Biochemical Pathology, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
  • Jessen N; Steno Diabetes Center Aarhus, Aarhus University Hospital, Aarhus, Denmark.
Am J Physiol Cell Physiol ; 321(2): C257-C268, 2021 08 01.
Article en En | MEDLINE | ID: mdl-34106790
Animal models clearly illustrate that the maintenance of skeletal muscle mass depends on the function and interaction of a heterogeneous population of resident and infiltrating mononuclear cells. Several lines of evidence suggest that mononuclear cells also play a role in muscle wasting in humans, and targeting these cells may open new treatment options for intervention or prevention in sarcopenia. Methodological and ethical constraints have perturbed exploration of the cellular characteristics and function of mononuclear cells in human skeletal muscle. Thus, investigations of cellular phenotypes often depend on immunohistochemical analysis of small tissue samples obtained by needle biopsies, which do not match the deep phenotyping of mononuclear cells obtained from animal models. Here, we have developed a protocol for fluorescence-activated cell sorting (FACS), based on single-cell RNA-sequencing data, for quantifying and characterizing mononuclear cell populations in human skeletal muscle. Muscle stem cells, fibro-adipogenic progenitors, and two subsets of macrophages (CD11c+/-) are present in needle biopsies in comparable quantities per milligram tissue to open surgical biopsies. We find that direct cell isolation is preferable due to a substantial shift in transcriptome when using preculture before the FACS procedure. Finally, in vitro validation of the cellular phenotype of muscle stem cells, fibro-adipogenic progenitors, and macrophages confirms population-specific traits. This study demonstrates that mononuclear cell populations can be quantified and subsequently analyzed from needle biopsy material and opens the perspective for future clinical studies of cellular mechanisms in muscle wasting.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Biopsia / Diferenciación Celular / Músculo Esquelético / Células Satélite del Músculo Esquelético Tipo de estudio: Guideline / Prognostic_studies Aspecto: Ethics Límite: Humans Idioma: En Revista: Am J Physiol Cell Physiol Asunto de la revista: FISIOLOGIA Año: 2021 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Biopsia / Diferenciación Celular / Músculo Esquelético / Células Satélite del Músculo Esquelético Tipo de estudio: Guideline / Prognostic_studies Aspecto: Ethics Límite: Humans Idioma: En Revista: Am J Physiol Cell Physiol Asunto de la revista: FISIOLOGIA Año: 2021 Tipo del documento: Article País de afiliación: Dinamarca Pais de publicación: Estados Unidos