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The term basal plate of the human placenta as a source of functional extravillous trophoblast cells
BACKGROUND: Extravillous trophoblast (EVT) cells are of pivotal importance in human embryo implantation and homeostasis of the maternal fetal interface. Invasion of the endometrium by EVT contributes to placental anchorage, spiral artery remodeling, immunological defense, tolerogenic responses, and...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3909387/ https://www.ncbi.nlm.nih.gov/pubmed/24467708 http://dx.doi.org/10.1186/1477-7827-12-7 |
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author | Borbely, Alexandre U Sandri, Silvana Fernandes, Isabella R Prado, Karen M Cardoso, Elaine C Correa-Silva, Simone Albuquerque, Renata Knöfler, Martin Beltrão-Braga, Patricia Campa, Ana Bevilacqua, Estela |
author_facet | Borbely, Alexandre U Sandri, Silvana Fernandes, Isabella R Prado, Karen M Cardoso, Elaine C Correa-Silva, Simone Albuquerque, Renata Knöfler, Martin Beltrão-Braga, Patricia Campa, Ana Bevilacqua, Estela |
author_sort | Borbely, Alexandre U |
collection | PubMed |
description | BACKGROUND: Extravillous trophoblast (EVT) cells are of pivotal importance in human embryo implantation and homeostasis of the maternal fetal interface. Invasion of the endometrium by EVT contributes to placental anchorage, spiral artery remodeling, immunological defense, tolerogenic responses, and several collaborative cross talks involved in establishing and maintaining a successful pregnancy. We report here an improved protocol for the isolation of fully differentiated EVT cells from the basal plate of the human term placenta. METHODS: The basal plate was carefully dissected from the villous tissue and the amniochorion membrane prior to enzymatic digestion. Term basal EVT cells were isolated using a 30 and 60% Percoll gradient. A panel of markers and characteristics of the isolated cells were used to confirm the specificity and efficiency of the method so that their potential as an investigative tool for placental research could be ascertained. RESULTS: Isolated cells were immunoreactive for cytokeratin-7 (CK-7), placental growth factor, placental alkaline phosphatase, human leukocyte antigen G1 (HLA-G1), and α1 and α5 integrins, similarly to the EVT markers from first trimester placental villi. Around 95% of the isolated cells labeled positively for CK-7 and 82% for HLA-G1. No significant change in viability was observed during 48 h of EVT culture as indicated by propidium iodide incorporation and trypan blue test exclusion. Genes for metalloproteinases MMP-2 and MMP9 (positive regulators of trophoblast invasiveness) were expressed up to 48 h of culturing, as also the gelatinolytic activity of the isolated cells. Transforming growth factor (TGF)-beta, which inhibits proliferation, migration, and invasiveness of first-trimester EVT cells, also reduced invasion of isolated term EVT cells in transwell assays, whereas epidermal growth factor was a positive modulator. CONCLUSIONS: Term basal plate may be a viable source of functional EVT cells that is an alternative to villous explant-derived EVT cells and cell lines. Isolated term EVT cells may be particularly useful in investigation of the role of trophoblast cells in pathological gestations, in which the precise regulation and interactive ability of extravillous trophoblast has been impaired. |
format | Online Article Text |
id | pubmed-3909387 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-39093872014-02-02 The term basal plate of the human placenta as a source of functional extravillous trophoblast cells Borbely, Alexandre U Sandri, Silvana Fernandes, Isabella R Prado, Karen M Cardoso, Elaine C Correa-Silva, Simone Albuquerque, Renata Knöfler, Martin Beltrão-Braga, Patricia Campa, Ana Bevilacqua, Estela Reprod Biol Endocrinol Research BACKGROUND: Extravillous trophoblast (EVT) cells are of pivotal importance in human embryo implantation and homeostasis of the maternal fetal interface. Invasion of the endometrium by EVT contributes to placental anchorage, spiral artery remodeling, immunological defense, tolerogenic responses, and several collaborative cross talks involved in establishing and maintaining a successful pregnancy. We report here an improved protocol for the isolation of fully differentiated EVT cells from the basal plate of the human term placenta. METHODS: The basal plate was carefully dissected from the villous tissue and the amniochorion membrane prior to enzymatic digestion. Term basal EVT cells were isolated using a 30 and 60% Percoll gradient. A panel of markers and characteristics of the isolated cells were used to confirm the specificity and efficiency of the method so that their potential as an investigative tool for placental research could be ascertained. RESULTS: Isolated cells were immunoreactive for cytokeratin-7 (CK-7), placental growth factor, placental alkaline phosphatase, human leukocyte antigen G1 (HLA-G1), and α1 and α5 integrins, similarly to the EVT markers from first trimester placental villi. Around 95% of the isolated cells labeled positively for CK-7 and 82% for HLA-G1. No significant change in viability was observed during 48 h of EVT culture as indicated by propidium iodide incorporation and trypan blue test exclusion. Genes for metalloproteinases MMP-2 and MMP9 (positive regulators of trophoblast invasiveness) were expressed up to 48 h of culturing, as also the gelatinolytic activity of the isolated cells. Transforming growth factor (TGF)-beta, which inhibits proliferation, migration, and invasiveness of first-trimester EVT cells, also reduced invasion of isolated term EVT cells in transwell assays, whereas epidermal growth factor was a positive modulator. CONCLUSIONS: Term basal plate may be a viable source of functional EVT cells that is an alternative to villous explant-derived EVT cells and cell lines. Isolated term EVT cells may be particularly useful in investigation of the role of trophoblast cells in pathological gestations, in which the precise regulation and interactive ability of extravillous trophoblast has been impaired. BioMed Central 2014-01-28 /pmc/articles/PMC3909387/ /pubmed/24467708 http://dx.doi.org/10.1186/1477-7827-12-7 Text en Copyright © 2014 Borbely et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Borbely, Alexandre U Sandri, Silvana Fernandes, Isabella R Prado, Karen M Cardoso, Elaine C Correa-Silva, Simone Albuquerque, Renata Knöfler, Martin Beltrão-Braga, Patricia Campa, Ana Bevilacqua, Estela The term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
title | The term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
title_full | The term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
title_fullStr | The term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
title_full_unstemmed | The term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
title_short | The term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
title_sort | term basal plate of the human placenta as a source of functional extravillous trophoblast cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3909387/ https://www.ncbi.nlm.nih.gov/pubmed/24467708 http://dx.doi.org/10.1186/1477-7827-12-7 |
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