title: Characterization of pluripotent stem cells.
abstract: Characterization of pluripotent stem cells is required for the registration of stem cell lines and allows for an impartial and objective comparison of the results obtained when generating multiple lines. It is therefore crucial to establish specific, fast and reliable protocols to detect the hallmarks of pluripotency. Such protocols should include immunocytochemistry (takes 2 d), identification of the three germ layers in in vitro-derived embryoid bodies by immunocytochemistry (immunodetection takes 3 d) and detection of differentiation markers in in vivo-generated teratomas by immunohistochemistry (differentiation marker detection takes 4 d). Standardization of the immunodetection protocols used ensures minimum variations owing to the source, the animal species, the endogenous fluorescence or the inability to collect large amounts of cells, thereby yielding results as fast as possible without loss of quality. This protocol provides a description of all the immunodetection procedures necessary to characterize mouse and human stem cell lines in different circumstances.
authors.affiliation: Center of Regenerative Medicine in Barcelona, Barcelona, Spain.
authors.forename: Mercè
authors.initials: M
authors.lastname: Martí
authors.forename: Lola
authors.initials: L
authors.lastname: Mulero
authors.forename: Cristina
authors.initials: C
authors.lastname: Pardo
authors.forename: Cristina
authors.initials: C
authors.lastname: Morera
authors.forename: Meritxell
authors.initials: M
authors.lastname: Carrió
authors.forename: Leopoldo
authors.initials: L
authors.lastname: Laricchia-Robbio
authors.forename: Concepcion Rodriguez
authors.initials: CR
authors.lastname: Esteban
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Izpisua Belmonte
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000592:standards*; D016172:DNA Fingerprinting; D058732:Embryoid Bodies / Q000166:cytology*; D005434:Flow Cytometry; D005855:Germ Layers / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007621:Karyotyping; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology*; D020033:Protein Isoforms / Q000378:metabolism; D013045:Species Specificity
chemical_list: D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D020033:Protein Isoforms