J 2019

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

BRAZERT, Maciej, Wieslawa KRANC, Blazej CHERMULA, Katarzyna KOWALSKA, Maurycy JANKOWSKI et. al.

Basic information

Original name

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

Authors

BRAZERT, Maciej (616 Poland), Wieslawa KRANC (616 Poland), Blazej CHERMULA (616 Poland), Katarzyna KOWALSKA (616 Poland), Maurycy JANKOWSKI (616 Poland), Piotr CELICHOWSKI (616 Poland), Michal JEŠETA (203 Czech Republic, belonging to the institution), Hanna PIOTROWSKA-KEMPISTY (616 Poland), Leszek PAWELCZYK (616 Poland), Maciej ZABEL (616 Poland), Paul MOZDZIAK (616 Poland) and Bartosz KEMPISTY (616 Poland, guarantor)

Edition

Journal of Clinical Medicine, BASEL, MDPI, 2019, 2077-0383

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30214 Obstetrics and gynaecology

Country of publisher

Switzerland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 3.303

RIV identification code

RIV/00216224:14110/19:00112837

Organization unit

Faculty of Medicine

DOI

UT WoS

000506640400002

Keywords in English

ovarian granulosa; human; in vitro; cell division

Tags

Tags

International impact, Reviewed
Změněno: 30/1/2020 13:41, Mgr. Tereza Miškechová

Abstract

V originále

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.