Endometrial stromal cells: isolation, expansion, morphological and functional properties

Aim: We aimed to study biological properties of human endometrial stromal cells in vitro. Materials and Methods: The endometrium samples (n = 5) were obtained by biopsy at the first phase of the menstrual cycle from women with endometrial hypoplasia. In all cases, a voluntary written informed consen...

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Published in:Experimental Oncology
Date:2017
Main Authors: Zlatska, A.V., Rodnichenko, A.E., Gubar, O.S., Zubov, D.O., Novikova, S.N., Vasyliev, R.G.
Format: Article
Language:English
Published: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2017
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Online Access:https://nasplib.isofts.kiev.ua/handle/123456789/138534
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Journal Title:Digital Library of Periodicals of National Academy of Sciences of Ukraine
Cite this:Endometrial stromal cells: isolation, expansion, morphological and functional properties / A.V. Zlatska, A.E. Rodnichenko, О.S. Gubar, D.О. Zubov, S.N. Novikova, R.G. Vasyliev // Experimental Oncology. — 2017 — Т. 39, № 3. — С. 197–202. — Бібліогр.: 12 назв. — англ.

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Digital Library of Periodicals of National Academy of Sciences of Ukraine
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author Zlatska, A.V.
Rodnichenko, A.E.
Gubar, O.S.
Zubov, D.O.
Novikova, S.N.
Vasyliev, R.G.
author_facet Zlatska, A.V.
Rodnichenko, A.E.
Gubar, O.S.
Zubov, D.O.
Novikova, S.N.
Vasyliev, R.G.
citation_txt Endometrial stromal cells: isolation, expansion, morphological and functional properties / A.V. Zlatska, A.E. Rodnichenko, О.S. Gubar, D.О. Zubov, S.N. Novikova, R.G. Vasyliev // Experimental Oncology. — 2017 — Т. 39, № 3. — С. 197–202. — Бібліогр.: 12 назв. — англ.
collection DSpace DC
container_title Experimental Oncology
description Aim: We aimed to study biological properties of human endometrial stromal cells in vitro. Materials and Methods: The endometrium samples (n = 5) were obtained by biopsy at the first phase of the menstrual cycle from women with endometrial hypoplasia. In all cases, a voluntary written informed consent was obtained from the patients. Endometrial fragments were dissociated by enzymatic treatment. The cells were cultured in DMEM/F12 supplemented with 10% FBS, 2 mМ L-glutamine and 1 ng/ml FGF-2 in a multi-gas incubator at 5% CO₂ and 5% O₂. At P3 the cells were subjected to immunophenotyping, multilineage differentiation, karyotype stability and colony forming efficiency. The cell secretome was assessed by BioRad Multiplex immunoassay kit. Results: Primary population of endometrial cells was heterogeneous and contained cells with fibroblast-like and epithelial-like morphology, but at P3 the majority of cell population had fibroblast-like morphology. The cells possessed typical for MSCs phenotype CD90⁺CD105⁺CD73⁺CD34⁻CD45⁻HLA⁻DR⁻. The cells also expressed CD140a, CD140b, CD146, and CD166 antigents; and were negative for CD106, CD184, CD271, and CD325. Cell doubling time was 29.6 ± 1.3 h. The cells were capable of directed osteogenic, adipogenic and chondrogenic differentiation. The cells showed 35.7% colony forming efficiency and a tendency to 3D spheroid formation. The GTG-banding assay confirmed the stability of eMSC karyotype during long-term culturing (up to P8). After 48 h incubation period in serum-free medium eMSC secreted anti-inflammatory IL-1ra, as well as IL-6, IL-8 and IFNγ, angiogenic factors VEGF, GM-CSF and FGF-2, chemokines IP-10 and MCP-1. Conclusion: Thus, cultured endometrial stromal cells meet minimal ISCT criteria for MSC. Proliferative potential, karyotype stability, multilineage plasticity and secretome profile make eMSC an attractive object for the regenerative medicine use.
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publishDate 2017
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
record_format dspace
spelling Zlatska, A.V.
Rodnichenko, A.E.
Gubar, O.S.
Zubov, D.O.
Novikova, S.N.
Vasyliev, R.G.
2018-06-19T09:06:09Z
2018-06-19T09:06:09Z
2017
Endometrial stromal cells: isolation, expansion, morphological and functional properties / A.V. Zlatska, A.E. Rodnichenko, О.S. Gubar, D.О. Zubov, S.N. Novikova, R.G. Vasyliev // Experimental Oncology. — 2017 — Т. 39, № 3. — С. 197–202. — Бібліогр.: 12 назв. — англ.
1812-9269
https://nasplib.isofts.kiev.ua/handle/123456789/138534
Aim: We aimed to study biological properties of human endometrial stromal cells in vitro. Materials and Methods: The endometrium samples (n = 5) were obtained by biopsy at the first phase of the menstrual cycle from women with endometrial hypoplasia. In all cases, a voluntary written informed consent was obtained from the patients. Endometrial fragments were dissociated by enzymatic treatment. The cells were cultured in DMEM/F12 supplemented with 10% FBS, 2 mМ L-glutamine and 1 ng/ml FGF-2 in a multi-gas incubator at 5% CO₂ and 5% O₂. At P3 the cells were subjected to immunophenotyping, multilineage differentiation, karyotype stability and colony forming efficiency. The cell secretome was assessed by BioRad Multiplex immunoassay kit. Results: Primary population of endometrial cells was heterogeneous and contained cells with fibroblast-like and epithelial-like morphology, but at P3 the majority of cell population had fibroblast-like morphology. The cells possessed typical for MSCs phenotype CD90⁺CD105⁺CD73⁺CD34⁻CD45⁻HLA⁻DR⁻. The cells also expressed CD140a, CD140b, CD146, and CD166 antigents; and were negative for CD106, CD184, CD271, and CD325. Cell doubling time was 29.6 ± 1.3 h. The cells were capable of directed osteogenic, adipogenic and chondrogenic differentiation. The cells showed 35.7% colony forming efficiency and a tendency to 3D spheroid formation. The GTG-banding assay confirmed the stability of eMSC karyotype during long-term culturing (up to P8). After 48 h incubation period in serum-free medium eMSC secreted anti-inflammatory IL-1ra, as well as IL-6, IL-8 and IFNγ, angiogenic factors VEGF, GM-CSF and FGF-2, chemokines IP-10 and MCP-1. Conclusion: Thus, cultured endometrial stromal cells meet minimal ISCT criteria for MSC. Proliferative potential, karyotype stability, multilineage plasticity and secretome profile make eMSC an attractive object for the regenerative medicine use.
en
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
Experimental Oncology
Original contributions
Endometrial stromal cells: isolation, expansion, morphological and functional properties
Article
published earlier
spellingShingle Endometrial stromal cells: isolation, expansion, morphological and functional properties
Zlatska, A.V.
Rodnichenko, A.E.
Gubar, O.S.
Zubov, D.O.
Novikova, S.N.
Vasyliev, R.G.
Original contributions
title Endometrial stromal cells: isolation, expansion, morphological and functional properties
title_full Endometrial stromal cells: isolation, expansion, morphological and functional properties
title_fullStr Endometrial stromal cells: isolation, expansion, morphological and functional properties
title_full_unstemmed Endometrial stromal cells: isolation, expansion, morphological and functional properties
title_short Endometrial stromal cells: isolation, expansion, morphological and functional properties
title_sort endometrial stromal cells: isolation, expansion, morphological and functional properties
topic Original contributions
topic_facet Original contributions
url https://nasplib.isofts.kiev.ua/handle/123456789/138534
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AT rodnichenkoae endometrialstromalcellsisolationexpansionmorphologicalandfunctionalproperties
AT gubaros endometrialstromalcellsisolationexpansionmorphologicalandfunctionalproperties
AT zubovdo endometrialstromalcellsisolationexpansionmorphologicalandfunctionalproperties
AT novikovasn endometrialstromalcellsisolationexpansionmorphologicalandfunctionalproperties
AT vasylievrg endometrialstromalcellsisolationexpansionmorphologicalandfunctionalproperties