Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages

Giulio Alessandri, Stefano Pagano, Alessandra Bez, Anna Benetti, Stefano Pozzi, Gioacchin Iannolo, Manuela Baronio, Gloria Invernici, Arnaldo Caruso, Claudio Muneretto, Gianluigi Bisleri, Eugenio Parati

Research output: Contribution to journalArticle

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Abstract

Background Skeletal-muscle-derived stem cells seem to be a distinct population of immature progenitors of satellite cells, but their functional properties remain unclear, especially in human adult tissue. We investigated their differentiation in samples of skeletal muscle obtained from adults undergoing cardiovascular surgery. Methods Samples were obtained from the brachioradialis muscle of 12 patients in whom the radial artery was the conduit for myocardial revascularisation. The stem cells were isolated by a procedure similar to that used for rat gastrocnemius and cultured in medium optimised for growth of neural stem cells. Cytometry was used for phenotypic characterisation and immunocytochemistry and RT-PCR to assess differentiation. Immunohistochemistry was used to examine engraftment of skeletal-muscle-derived stem cells into injured rat spinal cord. Findings The skeletal-muscle stem cells consisted of two distinct types: one with the typical spindle morphology of satellite cells, the other of rounded cells. Some cultures could be maintained for longer than 6 months. The cells were mainly positive for desmin and to a lesser extent CD105, vimentin, and AC133/CD133, but negative for FLK-1/KDR, CD34, CD31, CD45, von Willebrand factor, Ve-cadherins, and BCL2. After in-vitro differentiation, the cells were able to organise skeletal-muscle fibres and stained positively for striated-muscle actin, smooth-muscle actin, and desmin. Moreover, they differentiated into astrocytes and neurons, as confirmed by positive staining for characteristic proteins. Interpretation Adult human skeletal muscle includes a population of progenitor stem cells that can generate cells of the same lineage and cells with neurogenic properties. Muscle may therefore be a tissue source for the isolation of pluripotent stem cells for development of cell-based therapies for human myogenic and neurogenic diseases.

Original languageEnglish
Pages (from-to)1872-1883
Number of pages12
JournalLancet
Volume364
Issue number9448
DOIs
Publication statusPublished - Nov 20 2004

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Cell Lineage
Stem Cells
Muscles
Skeletal Muscle
Desmin
Actins
Immunohistochemistry
Pluripotent Stem Cells
Myocardial Revascularization
Radial Artery
Striated Muscle
Neural Stem Cells
Skeletal Muscle Fibers
von Willebrand Factor
Vimentin
Cadherins
Cell- and Tissue-Based Therapy
Astrocytes
Muscle Cells
Population

ASJC Scopus subject areas

  • Medicine(all)

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Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages. / Alessandri, Giulio; Pagano, Stefano; Bez, Alessandra; Benetti, Anna; Pozzi, Stefano; Iannolo, Gioacchin; Baronio, Manuela; Invernici, Gloria; Caruso, Arnaldo; Muneretto, Claudio; Bisleri, Gianluigi; Parati, Eugenio.

In: Lancet, Vol. 364, No. 9448, 20.11.2004, p. 1872-1883.

Research output: Contribution to journalArticle

Alessandri, G, Pagano, S, Bez, A, Benetti, A, Pozzi, S, Iannolo, G, Baronio, M, Invernici, G, Caruso, A, Muneretto, C, Bisleri, G & Parati, E 2004, 'Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages', Lancet, vol. 364, no. 9448, pp. 1872-1883. https://doi.org/10.1016/S0140-6736(04)17443-6
Alessandri, Giulio ; Pagano, Stefano ; Bez, Alessandra ; Benetti, Anna ; Pozzi, Stefano ; Iannolo, Gioacchin ; Baronio, Manuela ; Invernici, Gloria ; Caruso, Arnaldo ; Muneretto, Claudio ; Bisleri, Gianluigi ; Parati, Eugenio. / Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages. In: Lancet. 2004 ; Vol. 364, No. 9448. pp. 1872-1883.
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T1 - Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages

AU - Alessandri, Giulio

AU - Pagano, Stefano

AU - Bez, Alessandra

AU - Benetti, Anna

AU - Pozzi, Stefano

AU - Iannolo, Gioacchin

AU - Baronio, Manuela

AU - Invernici, Gloria

AU - Caruso, Arnaldo

AU - Muneretto, Claudio

AU - Bisleri, Gianluigi

AU - Parati, Eugenio

PY - 2004/11/20

Y1 - 2004/11/20

N2 - Background Skeletal-muscle-derived stem cells seem to be a distinct population of immature progenitors of satellite cells, but their functional properties remain unclear, especially in human adult tissue. We investigated their differentiation in samples of skeletal muscle obtained from adults undergoing cardiovascular surgery. Methods Samples were obtained from the brachioradialis muscle of 12 patients in whom the radial artery was the conduit for myocardial revascularisation. The stem cells were isolated by a procedure similar to that used for rat gastrocnemius and cultured in medium optimised for growth of neural stem cells. Cytometry was used for phenotypic characterisation and immunocytochemistry and RT-PCR to assess differentiation. Immunohistochemistry was used to examine engraftment of skeletal-muscle-derived stem cells into injured rat spinal cord. Findings The skeletal-muscle stem cells consisted of two distinct types: one with the typical spindle morphology of satellite cells, the other of rounded cells. Some cultures could be maintained for longer than 6 months. The cells were mainly positive for desmin and to a lesser extent CD105, vimentin, and AC133/CD133, but negative for FLK-1/KDR, CD34, CD31, CD45, von Willebrand factor, Ve-cadherins, and BCL2. After in-vitro differentiation, the cells were able to organise skeletal-muscle fibres and stained positively for striated-muscle actin, smooth-muscle actin, and desmin. Moreover, they differentiated into astrocytes and neurons, as confirmed by positive staining for characteristic proteins. Interpretation Adult human skeletal muscle includes a population of progenitor stem cells that can generate cells of the same lineage and cells with neurogenic properties. Muscle may therefore be a tissue source for the isolation of pluripotent stem cells for development of cell-based therapies for human myogenic and neurogenic diseases.

AB - Background Skeletal-muscle-derived stem cells seem to be a distinct population of immature progenitors of satellite cells, but their functional properties remain unclear, especially in human adult tissue. We investigated their differentiation in samples of skeletal muscle obtained from adults undergoing cardiovascular surgery. Methods Samples were obtained from the brachioradialis muscle of 12 patients in whom the radial artery was the conduit for myocardial revascularisation. The stem cells were isolated by a procedure similar to that used for rat gastrocnemius and cultured in medium optimised for growth of neural stem cells. Cytometry was used for phenotypic characterisation and immunocytochemistry and RT-PCR to assess differentiation. Immunohistochemistry was used to examine engraftment of skeletal-muscle-derived stem cells into injured rat spinal cord. Findings The skeletal-muscle stem cells consisted of two distinct types: one with the typical spindle morphology of satellite cells, the other of rounded cells. Some cultures could be maintained for longer than 6 months. The cells were mainly positive for desmin and to a lesser extent CD105, vimentin, and AC133/CD133, but negative for FLK-1/KDR, CD34, CD31, CD45, von Willebrand factor, Ve-cadherins, and BCL2. After in-vitro differentiation, the cells were able to organise skeletal-muscle fibres and stained positively for striated-muscle actin, smooth-muscle actin, and desmin. Moreover, they differentiated into astrocytes and neurons, as confirmed by positive staining for characteristic proteins. Interpretation Adult human skeletal muscle includes a population of progenitor stem cells that can generate cells of the same lineage and cells with neurogenic properties. Muscle may therefore be a tissue source for the isolation of pluripotent stem cells for development of cell-based therapies for human myogenic and neurogenic diseases.

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