Vol 84, No 12 (2013)
ARTICLES
Expression and co-expression of surface markers of pluripotency on human amniotic cells cultured in different growth media
Aleksandra Bryzek, Piotr Czekaj, Danuta Plewka, Marcin Tomsia, Halina Komarska, Marta Lesiak, Aleksander L. Sieroń, Jerzy Sikora, Katarzyna Kopaczka
DOI: 10.17772/gp/1673
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Ginekol Pol 2013;84(12).
Vol 84, No 12 (2013)
ARTICLES
Abstract
Objectives: 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. Aim: 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. Material and methods: Immunofluorescence and fluorescence microscopy were used to identify and localize S.C. 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. Results and 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 S.C. 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.
Abstract
Objectives: 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. Aim: 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. Material and methods: Immunofluorescence and fluorescence microscopy were used to identify and localize S.C. 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. Results and 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 S.C. 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.
Keywords
human placenta, Amnion, pulripotent stem cells, markers of pluripotency, coexpression, SSEA-3, SSEA-4, TRA-1-60
Title
Expression and co-expression of surface markers of pluripotency on human amniotic cells cultured in different growth media
Journal
Ginekologia Polska
Issue
Vol 84, No 12 (2013)
Page views
922
Article views/downloads
1123
DOI
10.17772/gp/1673
Bibliographic record
Ginekol Pol 2013;84(12).
Keywords
human placenta
Amnion
pulripotent stem cells
markers of pluripotency
coexpression
SSEA-3
SSEA-4
TRA-1-60
Authors
Aleksandra Bryzek
Piotr Czekaj
Danuta Plewka
Marcin Tomsia
Halina Komarska
Marta Lesiak
Aleksander L. Sieroń
Jerzy Sikora
Katarzyna Kopaczka
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