title: Cellular diversity within embryonic stem cells: pluripotent clonal sublines show distinct differentiation potential.
abstract: Embryonic stem cells (ESC), derived from the early inner cell mass (ICM), are constituted of theoretically homogeneous pluripotent cells. Our study was designed to test this concept using experimental approaches that allowed characterization of progenies derived from single parental mouse ESC. Flow cytometry analysis showed that a fraction of ESC submitted to neural differentiation generates progenies that escape the desired phenotype. Live imaging of individual cells demonstrated significant variations in the capacity of parental ESC to generate neurons, raising the possibility of clonal diversity among ESC. To further substantiate this hypothesis, clonal sublines from ESC were generated by limit dilution. Transcriptome analysis of undifferentiated sublines showed marked differences in gene expression despite the fact that all clones expressed pluripotency markers. Sublines showed distinct differentiation potential, both in phenotypic differentiation assays and with respect to gene expression in embryoid bodies. Clones generated from another ESC line also showed individualities in their differentiation potential, demonstrating the wider applicability of these findings. Taken together, our observations demonstrate that pluripotent ESC consist of individual cell types with distinct differentiation potentials. These findings identify novel elements for the biological understanding of ESC and provide new tools with a major potential for their future in vitro and in vivo use.
authors.affiliation: Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Geneva, Switzerland. yannick.martinez@unige.ch
authors.forename: Yannick
authors.initials: Y
authors.lastname: Martinez
authors.forename: Frédérique
authors.initials: F
authors.lastname: Béna
authors.forename: Stefania
authors.initials: S
authors.lastname: Gimelli
authors.forename: Diderik
authors.initials: D
authors.lastname: Tirefort
authors.forename: Michel
authors.initials: M
authors.lastname: Dubois-Dauphin
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Krause
authors.forename: Olivier
authors.initials: O
authors.lastname: Preynat-Seauve
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002460:Cell Line; D002999:Clone Cells; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental; D014644:Genetic Variation; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D059010:Single-Cell Analysis; D059467:Transcriptome
chemical_list: D015415:Biomarkers