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Isolation and characterization of muscle stem cells, fibro-adipogenic progenitors and macrophages from human skeletal muscle biopsies

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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 et al.

I: American journal of physiology. Cell physiology, Bind 321, Nr. 2, 08.2021, s. C257-C268.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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@article{5296d24baeed488cbd1d8a4e0bbeee10,
title = "Isolation and characterization of muscle stem cells, fibro-adipogenic progenitors and macrophages from human skeletal muscle biopsies",
abstract = "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 {\th} / –) 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. ",
keywords = "Biopsy, Mononuclear cells, Skeletal muscle",
author = "Jensen, {Jonas B} and M{\o}ller, {Andreas B} and Jesper Just and Maike Mose and {de Paoli}, {Frank V} and Billeskov, {Tine B} and Fred, {Rikard G} and Pers, {Tune H} and Pedersen, {Steen B} and Petersen, {Klaus K} and Mette Bjerre and Jean Farup and Niels Jessen",
year = "2021",
month = aug,
doi = "10.1152/ajpcell.00127.2021",
language = "English",
volume = "321",
pages = "C257--C268",
journal = "American Journal of Physiology: Cell Physiology",
issn = "0363-6143",
publisher = "American Physiological Society",
number = "2",

}

RIS

TY - JOUR

T1 - Isolation and characterization of muscle stem cells, fibro-adipogenic progenitors and macrophages from human skeletal muscle biopsies

AU - Jensen, Jonas B

AU - Møller, Andreas B

AU - Just, Jesper

AU - Mose, Maike

AU - de Paoli, Frank V

AU - Billeskov, Tine B

AU - Fred, Rikard G

AU - Pers, Tune H

AU - Pedersen, Steen B

AU - Petersen, Klaus K

AU - Bjerre, Mette

AU - Farup, Jean

AU - Jessen, Niels

PY - 2021/8

Y1 - 2021/8

N2 - 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 þ / –) 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.

AB - 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 þ / –) 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.

KW - Biopsy

KW - Mononuclear cells

KW - Skeletal muscle

U2 - 10.1152/ajpcell.00127.2021

DO - 10.1152/ajpcell.00127.2021

M3 - Journal article

C2 - 34106790

VL - 321

SP - C257-C268

JO - American Journal of Physiology: Cell Physiology

JF - American Journal of Physiology: Cell Physiology

SN - 0363-6143

IS - 2

ER -