title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons.
abstract: Human pluripotent stem cells (hPSC) have promise for regenerative medicine due to their auto-renovation and differentiation capacities. Nevertheless, there are several ethical and methodological issues about these cells that have not been resolved. Human amniotic epithelial cells (hAEC) have been proposed as source of pluripotent stem cells. Several groups have studied hAEC but have reported inconsistencies about their pluripotency properties. The aim of the present study was the in vitro characterization of hAEC collected from a Mexican population in order to identify transcription factors involved in the pluripotency circuitry and to determine their epigenetic state. Finally, we evaluated if these cells differentiate to cortical progenitors. We analyzed qualitatively and quantitatively the expression of the transcription factors of pluripotency (OCT4, SOX2, NANOG, KLF4 and REX1) by RT-PCR and RT-qPCR in hAEC. Also, we determined the presence of OCT4, SOX2, NANOG, SSEA3, SSEA4, TRA-1-60, E-cadherin, KLF4, TFE3 as well as the proliferation and epigenetic state by immunocytochemistry of the cells. Finally, hAEC were differentiated towards cortical progenitors using a protocol of two stages. Here we show that hAEC, obtained from a Mexican population and cultured in vitro (P0-P3), maintained the expression of several markers strongly involved in pluripotency maintenance (OCT4, SOX2, NANOG, TFE3, KLF4, SSEA3, SSEA4, TRA-1-60 and E-cadherin). Finally, when hAEC were treated with growth factors and small molecules, they expressed markers characteristic of cortical progenitors (TBR2, OTX2, NeuN and β-III-tubulin). Our results demonstrated that hAEC express naïve pluripotent markers (KLF4, REX1 and TFE3) as well as the cortical neuron phenotype after differentiation. This highlights the need for further investigation of hAEC as a possible source of hPSC.
authors.affiliation: Laboratorio de Citopatología Ambiental, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Campus Zacatenco, Unidad Profesional "Adolfo López Mateos", México D.F., México.|Departamento de Biología Celular, Instituto Nacional de Perinatología, Montes Urales 800, Col. Lomas Virreyes, CP 11000, México D.F., México.
authors.forename: Irma Lydia
authors.initials: IL
authors.lastname: García-Castro
authors.affiliation: Departamento de Biología Celular, Instituto Nacional de Perinatología, Montes Urales 800, Col. Lomas Virreyes, CP 11000, México D.F., México.
authors.forename: Guadalupe
authors.initials: G
authors.lastname: García-López
authors.affiliation: Departamento de Biología Celular, Instituto Nacional de Perinatología, Montes Urales 800, Col. Lomas Virreyes, CP 11000, México D.F., México.
authors.forename: Daniela
authors.initials: D
authors.lastname: Ávila-González
authors.affiliation: Departamento de Inmuno-Bioquímica, Instituto Nacional de Perinatología, Montes Urales 800, Col. Lomas Virreyes, CP 11000, México D.F., México.
authors.forename: Héctor
authors.initials: H
authors.lastname: Flores-Herrera
authors.affiliation: Departamento de Biología Celular, Instituto Nacional de Perinatología, Montes Urales 800, Col. Lomas Virreyes, CP 11000, México D.F., México.
authors.forename: Anayansi
authors.initials: A
authors.lastname: Molina-Hernández
authors.affiliation: Departamento de Neurobiología Conductal y Cognitiva, Instituto de Neurobiología, UNAM, Querétaro, México.
authors.forename: Wendy
authors.initials: W
authors.lastname: Portillo
authors.affiliation: Laboratorio de Citopatología Ambiental, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Campus Zacatenco, Unidad Profesional "Adolfo López Mateos", México D.F., México.
authors.forename: Eva
authors.initials: E
authors.lastname: Ramón-Gallegos
authors.affiliation: Departamento de Biología Celular, Instituto Nacional de Perinatología, Montes Urales 800, Col. Lomas Virreyes, CP 11000, México D.F., México.
authors.forename: Néstor Fabián
authors.initials: NF
authors.lastname: Díaz
mesh_terms: D000650:Amnion / Q000378:metabolism* / Q000502:physiology; D015415:Biomarkers / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D044127:Epigenesis, Genetic / Q000502:physiology; D004847:Epithelial Cells / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D009474:Neurons / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000378:metabolism* / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D015415:Biomarkers; D036341:Intercellular Signaling Peptides and Proteins; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors