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Derivation and Characterization of Immortalized Human Muscle Satellite Cell Clones from Muscular Dystrophy Patients and Healthy Individuals.
Cells. 2020 Jul 26; 9(8)C

Abstract

In Duchenne muscular dystrophy (DMD) patients, absence of dystrophin causes muscle wasting by impacting both the myofiber integrity and the properties of muscle stem cells (MuSCs). Investigation of DMD encompasses the use of MuSCs issued from human skeletal muscle. However, DMD-derived MuSC usage is restricted by the limited number of divisions that human MuSCs can undertake in vitro before losing their myogenic characteristics and by the scarcity of human material available from DMD muscle. To overcome these limitations, immortalization of MuSCs appears as a strategy. Here, we used CDK4/hTERT expression in primary MuSCs and we derived MuSC clones from a series of clinically and genetically characterized patients, including eight DMD patients with various mutations, four congenital muscular dystrophies and three age-matched control muscles. Immortalized cultures were sorted into single cells and expanded as clones into homogeneous populations. Myogenic characteristics and differentiation potential were tested for each clone. Finally, we screened various promoters to identify the preferred gene regulatory unit that should be used to ensure stable expression in the human MuSC clones. The 38 clonal immortalized myogenic cell clones provide a large collection of controls and DMD clones with various genetic defects and are available to the academic community.

Authors+Show Affiliations

Institut NeuroMyoGène, Université Claude Bernard Lyon 1, Université de Lyon, CNRS 5310, INSERM U1217, 69008 Lyon, France.Institut Cochin, INSERM U1016, CNRS UMR8104, Université Paris Descartes, Université Sorbonne Paris Cité, 75014 Paris, France.Institut Cochin, INSERM U1016, CNRS UMR8104, Université Paris Descartes, Université Sorbonne Paris Cité, 75014 Paris, France.Institut Cochin, INSERM U1016, CNRS UMR8104, Université Paris Descartes, Université Sorbonne Paris Cité, 75014 Paris, France.Laboratoire de Culture Cellulaire, Service de Génétique et Biologie Moléculaires-Hôpital Cochin, APHP.5, Assistance Publique-Hôpitaux de Paris, 75014 Paris, France.Laboratoire de Culture Cellulaire, Service de Génétique et Biologie Moléculaires-Hôpital Cochin, APHP.5, Assistance Publique-Hôpitaux de Paris, 75014 Paris, France.Sprott Center for Stem Cell Research, Ottawa Hospital Research Institute; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.Institut Cochin, INSERM U1016, CNRS UMR8104, Université Paris Descartes, Université Sorbonne Paris Cité, 75014 Paris, France.Institut NeuroMyoGène, Université Claude Bernard Lyon 1, Université de Lyon, CNRS 5310, INSERM U1217, 69008 Lyon, France.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32722643

Citation

Massenet, Jimmy, et al. "Derivation and Characterization of Immortalized Human Muscle Satellite Cell Clones From Muscular Dystrophy Patients and Healthy Individuals." Cells, vol. 9, no. 8, 2020.
Massenet J, Gitiaux C, Magnan M, et al. Derivation and Characterization of Immortalized Human Muscle Satellite Cell Clones from Muscular Dystrophy Patients and Healthy Individuals. Cells. 2020;9(8).
Massenet, J., Gitiaux, C., Magnan, M., Cuvellier, S., Hubas, A., Nusbaum, P., Dilworth, F. J., Desguerre, I., & Chazaud, B. (2020). Derivation and Characterization of Immortalized Human Muscle Satellite Cell Clones from Muscular Dystrophy Patients and Healthy Individuals. Cells, 9(8). https://doi.org/10.3390/cells9081780
Massenet J, et al. Derivation and Characterization of Immortalized Human Muscle Satellite Cell Clones From Muscular Dystrophy Patients and Healthy Individuals. Cells. 2020 Jul 26;9(8) PubMed PMID: 32722643.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Derivation and Characterization of Immortalized Human Muscle Satellite Cell Clones from Muscular Dystrophy Patients and Healthy Individuals. AU - Massenet,Jimmy, AU - Gitiaux,Cyril, AU - Magnan,Mélanie, AU - Cuvellier,Sylvain, AU - Hubas,Arnaud, AU - Nusbaum,Patrick, AU - Dilworth,F Jeffrey, AU - Desguerre,Isabelle, AU - Chazaud,Bénédicte, Y1 - 2020/07/26/ PY - 2020/06/29/received PY - 2020/07/17/revised PY - 2020/07/22/accepted PY - 2020/7/30/entrez PY - 2020/7/30/pubmed PY - 2020/7/30/medline KW - Duchenne muscular dystrophy KW - congenital myopathies KW - degenerative myopathies KW - human muscle stem cells KW - immortalization JF - Cells JO - Cells VL - 9 IS - 8 N2 - In Duchenne muscular dystrophy (DMD) patients, absence of dystrophin causes muscle wasting by impacting both the myofiber integrity and the properties of muscle stem cells (MuSCs). Investigation of DMD encompasses the use of MuSCs issued from human skeletal muscle. However, DMD-derived MuSC usage is restricted by the limited number of divisions that human MuSCs can undertake in vitro before losing their myogenic characteristics and by the scarcity of human material available from DMD muscle. To overcome these limitations, immortalization of MuSCs appears as a strategy. Here, we used CDK4/hTERT expression in primary MuSCs and we derived MuSC clones from a series of clinically and genetically characterized patients, including eight DMD patients with various mutations, four congenital muscular dystrophies and three age-matched control muscles. Immortalized cultures were sorted into single cells and expanded as clones into homogeneous populations. Myogenic characteristics and differentiation potential were tested for each clone. Finally, we screened various promoters to identify the preferred gene regulatory unit that should be used to ensure stable expression in the human MuSC clones. The 38 clonal immortalized myogenic cell clones provide a large collection of controls and DMD clones with various genetic defects and are available to the academic community. SN - 2073-4409 UR - https://www.unboundmedicine.com/medline/citation/32722643/Derivation_and_Characterization_of_Immortalized_Human_Muscle_Satellite_Cell_Clones_from_Muscular_Dystrophy_Patients_and_Healthy_Individuals_ L2 - https://www.mdpi.com/resolver?pii=cells9081780 DB - PRIME DP - Unbound Medicine ER -
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