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Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation

Granulosa cells (GCs) are a population of somatic cells whose role after ovulation is progesterone production. GCs were collected from patients undergoing controlled ovarian stimulation during an in vitro fertilization procedure, and they were maintained for 1, 7, 15, and 30 days of in vitro primary...

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Autores principales: Brązert, Maciej, Kranc, Wiesława, Chermuła, Błażej, Kowalska, Katarzyna, Jankowski, Maurycy, Celichowski, Piotr, Jeseta, Michal, Piotrowska-Kempisty, Hanna, Pawelczyk, Leszek, Zabel, Maciej, Mozdziak, Paul, Kempisty, Bartosz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6947147/
https://www.ncbi.nlm.nih.gov/pubmed/31756998
http://dx.doi.org/10.3390/jcm8122026
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author Brązert, Maciej
Kranc, Wiesława
Chermuła, Błażej
Kowalska, Katarzyna
Jankowski, Maurycy
Celichowski, Piotr
Jeseta, Michal
Piotrowska-Kempisty, Hanna
Pawelczyk, Leszek
Zabel, Maciej
Mozdziak, Paul
Kempisty, Bartosz
author_facet Brązert, Maciej
Kranc, Wiesława
Chermuła, Błażej
Kowalska, Katarzyna
Jankowski, Maurycy
Celichowski, Piotr
Jeseta, Michal
Piotrowska-Kempisty, Hanna
Pawelczyk, Leszek
Zabel, Maciej
Mozdziak, Paul
Kempisty, Bartosz
author_sort Brązert, Maciej
collection PubMed
description Granulosa cells (GCs) are a population of somatic cells whose role after ovulation is progesterone production. GCs were collected from patients undergoing controlled ovarian stimulation during an in vitro fertilization procedure, and they were maintained for 1, 7, 15, and 30 days of in vitro primary culture before collection for further gene expression analysis. A study of genes involved in the biological processes of interest was carried out using expression microarrays. To validate the obtained results, Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) was performed. The direction of changes in the expression of the selected genes was confirmed in most of the examples. Six ontological groups (“cell cycle arrest”, “cell cycle process”, “mitotic spindle organization”, “mitotic spindle assembly checkpoint”, “mitotic spindle assembly”, and “mitotic spindle checkpoint”) were analyzed in this study. The results of the microarrays obtained by us allowed us to identify two groups of genes whose expressions were the most upregulated (FAM64A, ANLN, TOP2A, CTGF, CEP55, BIRC5, PRC1, DLGAP5, GAS6, and NDRG1) and the most downregulated (EREG, PID1, INHA, RHOU, CXCL8, SEPT6, EPGN, RDX, WNT5A, and EZH2) during the culture. The cellular ultrastructure showed the presence of structures characteristic of mitotic cell division: a centrosome surrounded by a pericentric matrix, a microtubule system, and a mitotic spindle connected to chromosomes. The main goal of the study was to identify the genes involved in mitotic division and to identify the cellular ultrastructure of GCs in a long-term in vitro culture. All of the genes in these groups were subjected to downstream analysis, and their function and relation to the ovarian environment are discussed. The obtained results suggest that long-term in vitro cultivation of GCs may lead to their differentiation toward another cell type, including cells with cancer-like characteristics.
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spelling pubmed-69471472020-01-13 Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation Brązert, Maciej Kranc, Wiesława Chermuła, Błażej Kowalska, Katarzyna Jankowski, Maurycy Celichowski, Piotr Jeseta, Michal Piotrowska-Kempisty, Hanna Pawelczyk, Leszek Zabel, Maciej Mozdziak, Paul Kempisty, Bartosz J Clin Med Article Granulosa cells (GCs) are a population of somatic cells whose role after ovulation is progesterone production. GCs were collected from patients undergoing controlled ovarian stimulation during an in vitro fertilization procedure, and they were maintained for 1, 7, 15, and 30 days of in vitro primary culture before collection for further gene expression analysis. A study of genes involved in the biological processes of interest was carried out using expression microarrays. To validate the obtained results, Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) was performed. The direction of changes in the expression of the selected genes was confirmed in most of the examples. Six ontological groups (“cell cycle arrest”, “cell cycle process”, “mitotic spindle organization”, “mitotic spindle assembly checkpoint”, “mitotic spindle assembly”, and “mitotic spindle checkpoint”) were analyzed in this study. The results of the microarrays obtained by us allowed us to identify two groups of genes whose expressions were the most upregulated (FAM64A, ANLN, TOP2A, CTGF, CEP55, BIRC5, PRC1, DLGAP5, GAS6, and NDRG1) and the most downregulated (EREG, PID1, INHA, RHOU, CXCL8, SEPT6, EPGN, RDX, WNT5A, and EZH2) during the culture. The cellular ultrastructure showed the presence of structures characteristic of mitotic cell division: a centrosome surrounded by a pericentric matrix, a microtubule system, and a mitotic spindle connected to chromosomes. The main goal of the study was to identify the genes involved in mitotic division and to identify the cellular ultrastructure of GCs in a long-term in vitro culture. All of the genes in these groups were subjected to downstream analysis, and their function and relation to the ovarian environment are discussed. The obtained results suggest that long-term in vitro cultivation of GCs may lead to their differentiation toward another cell type, including cells with cancer-like characteristics. MDPI 2019-11-20 /pmc/articles/PMC6947147/ /pubmed/31756998 http://dx.doi.org/10.3390/jcm8122026 Text en © 2019 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Brązert, Maciej
Kranc, Wiesława
Chermuła, Błażej
Kowalska, Katarzyna
Jankowski, Maurycy
Celichowski, Piotr
Jeseta, Michal
Piotrowska-Kempisty, Hanna
Pawelczyk, Leszek
Zabel, Maciej
Mozdziak, Paul
Kempisty, Bartosz
Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation
title Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation
title_full Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation
title_fullStr Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation
title_full_unstemmed Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation
title_short Human Ovarian Granulosa Cells Isolated during an IVF Procedure Exhibit Differential Expression of Genes Regulating Cell Division and Mitotic Spindle Formation
title_sort human ovarian granulosa cells isolated during an ivf procedure exhibit differential expression of genes regulating cell division and mitotic spindle formation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6947147/
https://www.ncbi.nlm.nih.gov/pubmed/31756998
http://dx.doi.org/10.3390/jcm8122026
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