title: Expression and co-expression of surface markers of pluripotency on human amniotic cells cultured in different growth media.
abstract: OBJECTIVE
  Despite constant advances in the field of biology and medical application of human embryonic stem cells, the molecular mechanism of pluripotency remains largely unknown. So far, definitions of pluripotent stem cells (SC) have been based on a limited number of antigenic markers and have not allowed for unambiguous determination of the homogeneity of each subpopulation. Moreover, the use of some crucial pluripotency markers such as SSEA-3 and SSEA-4 has recently been questioned due to the possibility that the pattern of surface glycans may be changed depending on the content of the cell culture medium.
  
  
  OBJECTIVE
  Quantitative analysis of amniotic SC subpopulations cultured in different media, based on the following pluripotency surface markers: SSEA-3, SSEA-4, TRA- 1-60 and TRA- 1-81 expression and co-expression.
  
  
  METHODS
  Immunofluorescence and fluorescence microscopy were used to identify and localize SC within a normal human placenta at term. The number of SSEA-4+, SSEA-3+, TRA-1-60+ and TRA-1-81+ cells and cells with co-expression of the above mentioned markers, cultured in media containing different protein supplements of animal origin, was counted by flow cytometry
  
  
  CONCLUSIONS
  Cells with characteristics of embryonic SC were identified in the amniotic epithelium and the chorion, but not in the decidua basalis. Amniotic epithelium contained various types of SC, with SSEA-4+ as the most numerous. Disproportion in the number of SSEA-4+, SSEA-3+, TRA-1-60+ and TRA-1-81+ cells and cells characterized by co-expression of these antigens, as well as lack of quantitative differences between SC subpopulations cultured in different media, was observed. In conclusion, the amniotic epithelium is composed of SC at different stages of the development but human amnion might become an alternative source of SSEA-4+ embryonic-like SC. The composition of the evaluated media, characterized by different content of animal-derived proteins, does not influence the number of cells identified within the SC subpopulations.
authors: Department of Cytophysiology, Medical University of Silesia, Katowice, Poland.
authors: Aleksandra
authors: A
authors: Bryzek
authors: Department of Cytophysiology, Medical University of Silesia, Katowice, Poland.
authors: Piotr
authors: P
authors: Czekaj
authors: Department of Cytophysiology, Medical University of Silesia, Katowice, Poland.
authors: Danuta
authors: D
authors: Plewka
authors: Department of Molecular Biology, Medical University of Silesia, Katowice, Poland.
authors: Halina
authors: H
authors: Komarska
authors: Department of Cytophysiology, Medical University of Silesia, Katowice, Poland.
authors: Marcin
authors: M
authors: Tomsia
authors: Department of Molecular Biology, Medical University of Silesia, Katowice, Poland.
authors: Marta
authors: M
authors: Lesiak
authors: Department of Molecular Biology, Medical University of Silesia, Katowice, Poland.
authors: Aleksander L
authors: AL
authors: Sieroń
authors: Department of Perinatology and Gynecology, Central Clinical Hospital, Medical University of Silesia, Katowice, Poland.
authors: Jerzy
authors: J
authors: Sikora
authors: Students Scientific Society, Medical University of Silesia, Katowice, Poland.
authors: Katarzyna
authors: K
authors: Kopaczka
mesh_terms: D000293:Adolescent; D000328:Adult; D000653:Amniotic Fluid / Q000166:cytology* / Q000276:immunology*; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000032:analysis*; D015415:Biomarkers / Q000032:analysis; D002823:Chorion / Q000166:cytology / Q000276:immunology; D003470:Culture Media; D003656:Decidua / Q000166:cytology / Q000276:immunology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000276:immunology; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology; D005260:Female; D006801:Humans; D010920:Placenta / Q000166:cytology / Q000276:immunology; D039904:Pluripotent Stem Cells / Q000737:chemistry* / Q000166:cytology / Q000276:immunology*; D011247:Pregnancy; D011509:Proteoglycans / Q000032:analysis; D055746:Stage-Specific Embryonic Antigens / Q000032:analysis*; D055815:Young Adult
chemical_list: D000954:Antigens, Surface; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; D003470:Culture Media; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; C107064:TRA-1-60 antigen, human; C519803:TRA-1-81 antigen, human; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4