Michał M. Hryciuk , Filip Schröter , Luise Hennicke , Beate C. Braun
{"title":"Spheroid formation and luteinization of granulosa cells of felids in a long-term 3D culture","authors":"Michał M. Hryciuk , Filip Schröter , Luise Hennicke , Beate C. Braun","doi":"10.1016/j.diff.2023.03.002","DOIUrl":null,"url":null,"abstract":"<div><p>In the present study, granulosa cells (GCs) from domestic cats and Persian leopard were cultured and characterized from selected days. The culture period was divided into two phases: maintenance, which lasted for 7 days, and luteinization, which followed for up to 11 days. Luteinization was performed on ultra-low attachment plates, supporting the formation of spheroids in a medium supplemented with insulin, forskolin, and luteinizing hormone (LH). GCs of domestic cat produced estradiol (E2) and progesterone (P4) during the maintenance phase. The gene expressions of some proteins involved in steroidogenesis were stable (<em>STAR</em>, <em>HSD3B1</em>) or decreased over time (<em>CYP11A1</em>, <em>HSD17B1</em>, <em>CYP17A1</em>, and <em>CYP19A1</em>), which was similar to the expressions of gonatropin receptors (<em>LHCGR</em> and <em>FSHR</em>). During the luteinization phase, P4 concentration significantly increased (<em>P</em> < 0.05), and E2, in contrast to the proliferation phase, was below detection range. The expression of genes of proteins involved in steroidogenesis (<em>STAR</em>, <em>CYP11A1</em>, <em>HSD3B1</em>, <em>HSD17B1</em>, <em>CYP17A1</em>, and <em>CYP19A1</em>) and of gonadotropin receptors (<em>LHCGR</em> and <em>FSHR</em>) significantly increased during the luteinization period, but some expressions exhibited a decrease at the end of the phase (<em>LHCGR</em>, <em>FSHR</em>, <em>HSD17B1</em>, <em>CYP19A1</em>). The morphology of the luteinized GCs of domestic cat resembled large luteal cells and had numerous vacuole-like structures. Also, the GCs of Persian leopard underwent luteinization, shown by increasing P4 production and <em>HSD3B1</em> expression. This study confirms that GCs from felids can be luteinized in a 3D spheroid system which can be a basis for further studies on luteal cell function of felids. Additionally, we could show that the domestic cat can serve as a model species for establishing cell culture methods which can be transferred to other felids.</p></div>","PeriodicalId":50579,"journal":{"name":"Differentiation","volume":null,"pages":null},"PeriodicalIF":2.2000,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Differentiation","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S030146812300021X","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
In the present study, granulosa cells (GCs) from domestic cats and Persian leopard were cultured and characterized from selected days. The culture period was divided into two phases: maintenance, which lasted for 7 days, and luteinization, which followed for up to 11 days. Luteinization was performed on ultra-low attachment plates, supporting the formation of spheroids in a medium supplemented with insulin, forskolin, and luteinizing hormone (LH). GCs of domestic cat produced estradiol (E2) and progesterone (P4) during the maintenance phase. The gene expressions of some proteins involved in steroidogenesis were stable (STAR, HSD3B1) or decreased over time (CYP11A1, HSD17B1, CYP17A1, and CYP19A1), which was similar to the expressions of gonatropin receptors (LHCGR and FSHR). During the luteinization phase, P4 concentration significantly increased (P < 0.05), and E2, in contrast to the proliferation phase, was below detection range. The expression of genes of proteins involved in steroidogenesis (STAR, CYP11A1, HSD3B1, HSD17B1, CYP17A1, and CYP19A1) and of gonadotropin receptors (LHCGR and FSHR) significantly increased during the luteinization period, but some expressions exhibited a decrease at the end of the phase (LHCGR, FSHR, HSD17B1, CYP19A1). The morphology of the luteinized GCs of domestic cat resembled large luteal cells and had numerous vacuole-like structures. Also, the GCs of Persian leopard underwent luteinization, shown by increasing P4 production and HSD3B1 expression. This study confirms that GCs from felids can be luteinized in a 3D spheroid system which can be a basis for further studies on luteal cell function of felids. Additionally, we could show that the domestic cat can serve as a model species for establishing cell culture methods which can be transferred to other felids.
期刊介绍:
Differentiation is a multidisciplinary journal dealing with topics relating to cell differentiation, development, cellular structure and function, and cancer. Differentiation of eukaryotes at the molecular level and the use of transgenic and targeted mutagenesis approaches to problems of differentiation are of particular interest to the journal.
The journal will publish full-length articles containing original work in any of these areas. We will also publish reviews and commentaries on topics of current interest.
The principal subject areas the journal covers are: • embryonic patterning and organogenesis
• human development and congenital malformation
• mechanisms of cell lineage commitment
• tissue homeostasis and oncogenic transformation
• establishment of cellular polarity
• stem cell differentiation
• cell reprogramming mechanisms
• stability of the differentiated state
• cell and tissue interactions in vivo and in vitro
• signal transduction pathways in development and differentiation
• carcinogenesis and cancer
• mechanisms involved in cell growth and division especially relating to cancer
• differentiation in regeneration and ageing
• therapeutic applications of differentiation processes.