title: Distinct population of hair cell progenitors can be isolated from the postnatal mouse cochlea using side population analysis.
abstract: In mammals, the permanence of hearing loss is due mostly to the incapacity of the cochlea to replace lost mechano-receptor cells (i.e., hair cells [HCs]). The generation of new HCs from a renewable source of progenitors is a principal requirement for developing a cell therapy within this sensory organ. A subset of stem cells, termed side population (SP), has been identified in several tissues of mammals. The ATP-binding cassette transporter Abcg2/Bcrp1 contributes to the specification of the SP phenotype and is proposed as a universal marker for stem/progenitor cells. A defining character of these SP cells is a high efflux capacity for Hoechst dye. Here, we demonstrate that Abcg2 transporter is expressed with two other stem/progenitor cell markers (i.e., Nestin and Musashi1) in distinct and overlapping domains of the supporting cells within the postnatal cochlea. We have developed and describe a fluorescence-activated cell sorting (FACS) technique that enables the purification of a discrete subpopulation of SP-supporting cells from the early postnatal mouse cochlea based on their ability to exclude Hoechst dye. These FACS-isolated cells can divide and express markers of stem/progenitor cells such as Abcg2, a determinant of the SP phenotype, and Musashi1, a neural stem/progenitor cell marker. These markers can differentiate cells expressing markers of HCs and supporting cells in vitro. Our observation that these SP cells are capable of differentiating into HC-like cells implies a possible use for such cells (i.e., the replacement of lost auditory HCs within damaged cochlea).
authors.affiliation: Institute of Neuroscience, INSERM U.583, Montpellier, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Savary
authors.forename: Jean Philippe
authors.initials: JP
authors.lastname: Hugnot
authors.forename: Yolaine
authors.initials: Y
authors.lastname: Chassigneux
authors.forename: Cecile
authors.initials: C
authors.lastname: Travo
authors.forename: Christophe
authors.initials: C
authors.lastname: Duperray
authors.forename: Thomas
authors.initials: T
authors.lastname: Van De Water
authors.forename: Azel
authors.initials: A
authors.lastname: Zine
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers; D001973:Bromodeoxyuridine; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D003051:Cochlea / Q000166:cytology*; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000378:metabolism; D004398:Dyneins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006198:Hair Cells, Auditory / Q000166:cytology*; D007381:Intermediate Filament Proteins / Q000378:metabolism; D007760:Labyrinth Supporting Cells / Q000166:cytology; D051379:Mice; D000081412:Myosin VIIa; D009218:Myosins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D014018:Tissue Distribution
chemical_list: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; C501412:Abcg2 protein, mouse; D015415:Biomarkers; D007381:Intermediate Filament Proteins; C104786:Msi1h protein, mouse; C000631456:Myo7a protein, mouse; D000081412:Myosin VIIa; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D009218:Myosins; D004398:Dyneins; D001973:Bromodeoxyuridine