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Progenitor Cell Isolation From Muscle-derived Cells Based on Adhesion Properties

Karl Rouger¹, Benoît Fornasari¹, Valerie Armengol, Gregory Jouvion, Isabelle Leroux, Laurence Dubreil, Marie Feron, Laetitia Guevel and Yan Cherel

INRA, UMR703, Développement et Pathologie du Tissu Musculaire, Ecole Nationale Vétérinaire, Nantes, France (KR,BF,VA,GJ,IL,LD,YC); INSERM, UMR791, Laboratoire d'Ingenierie Ostéo-articulaire et Dentaire, Faculté de Chirurgie Dentaire, Nantes, France (VA); and Biotechnologie, Biocatalyse et Biorégulation (U3B), Faculté des Sciences et des Techniques, CNRS, UMR6204, Nantes, France (MF,LG)

Correspondence to: Karl Rouger, INRA, UMR703, Ecole Nationale Vétérinaire de Nantes, Route de Gachet, BP-40706, Nantes, F-44307 France. E-mail: rouger{at}vet-nantes.fr

Adult skeletal muscle possesses remarkable regenerative capacity that has conventionally been attributed to the satellite cells. These precursor cells were thought to contain distinct populations with varying myogenic potential. Recently, the identification of multipotent stem cells capable of new myofiber formation has expanded the general view on the muscle regenerative process. Here we examined the characteristics of turkey skeletal muscle-derived cell (MDC) populations that were separated according to their adhesion abilities. We sought to determine whether these abilities could be a potential tool for separating cells with different myogenic commitment. Using the preplate technique, we showed that MDCs display a wide range of adhesion ability, allowing us to isolate a marginal fraction with initial adhesion defect. Methodological investigations revealed that this defect represents an intrinsic and well-established biological feature for these cells. In vitro behavioral and morphological analyses showed that late adherent cells (LACs) share several primitive cell characteristics. Phenotypic assessment indicated that LACs contain early stage myogenic cells and immature progenitors of satellite cells, whereas early adherent cells consist mainly of fully committed precursors. Overall, our findings demonstrate for the first time in an avian model that differential MDC adhesion properties could be used to efficiently purify cells with varying myogenic commitment, including immature progenitor cells. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. (J Histochem Cytochem 55:607–618, 2007)

Key Words: myogenesis • muscle-derived cell • skeletal muscle • adhesion • preplate technique • avian

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