Expression of genes involved in the inflammatory response in human granulosa cells in short-term in vitro culture
Author:
Kałużna Sandra1, Bryl Rut2, Chermuła Błażej2, Sibiak Rafał1, Stefańska Katarzyna1, Pieńkowski Wojciech3, Kranc Wiesława4, Jeseta Michal5, Ventruba Pavel5, Zakova Jana5, Crha Igor5, Pawelczyk Leszek2, Spaczyński Robert Z.2
Affiliation:
1. Department of Histology and Embryology, Poznań University of Medical Sciences , Poznań , Poland 2. Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznań University of Medical Sciences , Poznań , Poland 3. Division of Perinatology and Women’s Diseases, Poznań University of Medical Sciences , Poznań , Poland 4. Department of Anatomy, Poznań University of Medical Sciences , Poznań , Poland 5. Department of Obstetrics and Gynecology, University Hospital and Masaryk University , Brno , Czech Republic
Abstract
Abstract
The essential function of granulosa cells is to maintain the proper course of oogenesis and folliculogenesis.
The immune system is an additional local regulator of ovarian function, with cytokines necessary for the proper function of the ovaries, including the secretion of steroid hormones
This study aimed to analyze the expression of genes in human GCs in short-term primary culture and define the difference in the expression of IL1β, IL6, and TNFα genes at 48h and 72h of culture compared to the 24h control. Total RNA was isolated using the Chomczyński and Sacchi protocol. RNA samples were treated with DNase I and reverse transcribed (RT) into cDNA. The determination of transcript levels of the mentioned genes was performed using the Light Cycler® 96 Real-Time PCR kit, Roche Diagnostics GmbH (Mannheim, Germany).
The present study proved that granulosa cells in a short-term primary in vitro culture express IL-1β, IL-6, and TNFα. The tested genes show a decrease in expression at 24h of culture and a subsequent slight increase at 72h, not exceeding the initial levels. The expression changes the most for IL1β and the least for TNFα.
The fluctuations in the amount of transcript may be influenced by factors stored in granulosa cells before the IVM procedure, the procedure of in vitro fertilization, as well as factors related to the process of primary culture. More research is needed to understand the details of these occurrences.
Running title: The inflammatory response in human granulosa cells
Publisher
Walter de Gruyter GmbH
Subject
Cell Biology,Molecular Biology
Reference34 articles.
1. Kulus M, Kranc W, Sujka-Kordowska P, Mozdziak P, Jankowski M, Konwerska A, Kulus J, Bukowska D, Skowroński M, Piotrowska-Kempisty H, Nowicki M, Kempisty B, Antosik P. The processes of cellular growth, aging, and programmed cell death are involved in lifespan of ovarian granulosa cells during short-term IVC – Study based on animal model. Theriogenology. 2020;148:76–88; DOI:10.1016/j.theriogenology.2020.02.044. 2. Dompe C, Kranc W, Jopek K, Kowalska K, Ciesiółka S, Chermuła B, Bryja A, Jankowski M, Perek J, Józkowiak M, Moncrieff L, Hutchings G, Janowicz K, Pawelczyk L, Bruska M, Petitte J, Mozdziak P, Kulus M, Piotrowska-Kempisty H, Spaczyński R, Nowicki M, Kempisty B. Muscle Cell Morphogenesis, Structure, Development and Differentiation Processes Are Significantly Regulated during Human Ovarian Granulosa Cells In Vitro Cultivation. J Clin Med. 2020;9:2006; DOI:10.3390/jcm9062006. 3. Brązert M, Kranc W, Celichowski P, Jankowski M, Piotrowska-Kempisty H, Pawelczyk L, Bruska M, Zabel M, Nowicki M, Kempisty B. Expression of genes involved in neurogenesis, and neuronal precursor cell proliferation and development: Novel pathways of human ovarian granulosa cell differentiation and transdifferentiation capability in vitro. Mol Med Rep. 2020;21:1749–60; DOI:10.3892/mmr.2020.10972. 4. Brązert M, Kranc W, Celichowski P, Ożegowska K, Budna-Tukan J, Jeseta M, Pawelczyk L, Bruska M, Zabel M, Nowicki M, Kempisty B. Novel markers of human ovarian granulosa cell differentiation toward osteoblast lineage: A microarray approach. Mol Med Rep. 2019;20:4403–14; DOI:10.3892/mmr.2019.10709. 5. Karagouni EE, Chryssikopoulos A, Mantzavinos T, Kanakas N, Dotsika EN. Interleukin-1β and interleukin-1α may affect the implantation rate of patients undergoing in vitro fertilization-embryo transfer. Fertil Steril. 1998;70:553–9; DOI:10.1016/S0015-0282(98)00243-X.
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