Establishment and validation of a model for non-luteinized human mural granulosa cell culture

• Published in: Molecular and cellular endocrinology Vol. 384; no. 1-2; pp. 165 - 174
• Main Authors: Ophir, L., Yung, Y., Maman, E., Rubinstein, N., Yerushalmi, G.M., Haas, J., Barzilay, E., Hourvitz, A.
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier Ireland Ltd 25.03.2014
• Subjects: Adult; Aromatase - genetics; Aromatase - metabolism; Biomarkers - metabolism; Cell culture; Cells, Cultured; Cholesterol Side-Chain Cleavage Enzyme - genetics; Cholesterol Side-Chain Cleavage Enzyme - metabolism; Chorionic Gonadotropin - pharmacology; Female; Follicle; Follicle Stimulating Hormone - pharmacology; Gene Expression; Granulosa Cells - cytology; Granulosa Cells - drug effects; Granulosa Cells - metabolism; Humans; Luteinization; Luteinization - genetics; Models, Biological; Mural granulosa cells; Ovary; Receptors, LHRH - genetics; Receptors, LHRH - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Follicle; Ovary; Cell culture; Luteinization; Mural granulosa cells
• ISSN: 0303-7207; 1872-8057
• DOI: 10.1016/j.mce.2014.01.018

•Cell culture techniques of human mural granulosa cells (MGCs) serve as a major in vitro tool.•We establish a standardized protocol for the culture of MGCs as a model for different stages of folliculogenesis.•Four days incubation of luteinized MGCs achieve genes expression levels similar to those of early non-luteinized follicles.•FSH stimulation for 48h showed ovulatory genes expression similar to the pre-ovulatory non-luteinized follicles.•Stimulated cells responded to hCG stimulation with pattern similar to the response of pre-ovulatory follicles. Cell culture techniques of human mural granulosa cells (MGCs) serve as a major in vitro tool. However, the use of luteinized MGCs has major limitations due to their luteinized state. Our aim was to establish a standardized protocol for the culture of MGCs as a model for different stages of folliculogenesis. We showed that early-non-luteinized, preovulatory-non-luteinized and luteal-MGCs have distinct gene expression pattern. After 4days of incubation of luteinized-MGCs, ovulatory genes mRNA’s achieve expression levels similar to the early non-luteinized follicles. FSH stimulation for 48h of these 4days cultured MGCs showed ovulatory genes mRNA’s expression similar to the pre-ovulatory non-luteinized follicles. These FSH-stimulated cells responded to hCG stimulation in a pattern similar to the response of pre-ovulatory follicles. This novel model may provide a standardized research tool for delineation of the molecular processes occurring during the latter stages of follicular development in the human ovary.