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Characterization of distinct mesenchymal-like cell populations from human skeletal muscle in situ and in vitro
Authors:Séverine Lecourt  Jean-Pierre Marolleau  Olivia Fromigué  Rina Andriamanalijaona  Jean-Thomas Vilquin
Institution:a UPMC/AIM UMR S 974, Groupe Hospitalier Pitié-Salpêtrière, Paris, France
b INSERM U974, Groupe Hospitalier Pitié-Salpêtrière, Paris, France
c CNRS UMR 7215, Groupe Hospitalier Pitié-Salpêtrière, Paris, France
d Laboratoire de Thérapie Cellulaire, Hôpital Saint Louis, Paris, France
e CHU Amiens Hôpital Sud, Service d'Hématologie Clinique, UPJV, Amiens, France
f INSERM U606, Université Paris 07, Hôpital Lariboisière, Paris, France
g Genzyme S.A.S., Saint-Germain en Laye, France
h Laboratoire de Biochimie des Tissus Conjonctifs, Faculté de Médecine, Caen, France
i Myosix S.A., Saint-Germain en Laye, France
Abstract:Human skeletal muscle is an essential source of various cellular progenitors with potential therapeutic perspectives. We first used extracellular markers to identify in situ the main cell types located in a satellite position or in the endomysium of the skeletal muscle. Immunohistology revealed labeling of cells by markers of mesenchymal (CD13, CD29, CD44, CD47, CD49, CD62, CD73, CD90, CD105, CD146, and CD15 in this study), myogenic (CD56), angiogenic (CD31, CD34, CD106, CD146), hematopoietic (CD10, CD15, CD34) lineages. We then analysed cell phenotypes and fates in short- and long-term cultures of dissociated muscle biopsies in a proliferation medium favouring the expansion of myogenic cells. While CD56+ cells grew rapidly, a population of CD15+ cells emerged, partly from CD56+ cells, and became individualized. Both populations expressed mesenchymal markers similar to that harboured by human bone marrow-derived mesenchymal stem cells. In differentiation media, both CD56+ and CD15+ cells shared osteogenic and chondrogenic abilities, while CD56+ cells presented a myogenic capacity and CD15+ cells presented an adipogenic capacity. An important proportion of cells expressed the CD34 antigen in situ and immediately after muscle dissociation. However, CD34 antigen did not persist in culture and this initial population gave rise to adipogenic cells. These results underline the diversity of human muscle cells, and the shared or restricted commitment abilities of the main lineages under defined conditions.
Keywords:ALP  Alkaline Phosphatase  BM  Bone Marrow  BM-MSC  Bone-arrow-derived mesenchymal stem cells  DFBS  Defined Foetal Bovine Serum  mAb  Monoclonal Antibody  MSC  Mesenchymal Stem Cell  αSMA  α Smooth Muscle Actin
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