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Extraction, Labeling, and Purification of Lineage-Specific Cells from Human Antral Follicles
Published on: November 30, 2022
What we have learned from isolated cells from human ovary?
Hitoshi Okamura1, Hidetaka Katabuchi, Takashi Ohba
1Department of Obstetrics and Gynecology, Kumamoto University School of Medicine, Honjo 1-1-1, Kumamoto 860-8556, Japan. hokamura@kaiju.medic.kumamoto-u.ac.jp
Molecular and Cellular Endocrinology
|May 29, 2003
Summary
Researchers developed immortalized human ovarian surface epithelium (OSE) and granulosa cells for studying ovarian function and disorders. These cell lines, particularly OSE, show potential links to endometriosis and steroidogenesis, while granulosa cells demonstrate SF-1-dependent steroid production.
Area of Science:
- Reproductive biology and cell biology
- Gynecologic pathology and oncology
Background:
- Ovarian function involves complex morphodynamics and tropic factor control, typically studied in animals due to limited human specimens.
- Endometriosis may arise from coelomic metaplasia of the peritoneal mesothelium, including ovarian surface epithelium (OSE).
- Understanding granulosa cell molecular pathways is crucial for follicle maturation but remains largely unknown.
Purpose of the Study:
- To establish and characterize immortalized human ovarian surface epithelium (OSE) and granulosa cell lines for advanced research.
- To investigate the potential of OSE in endometriosis development and steroidogenic capabilities.
- To elucidate the role of Steroidogenic Factor-1 (SF-1) in human granulosa cell steroidogenesis.
Main Methods:
- Development of in vitro culture and immortalization systems for human OSE and granulosa cells.
- Co-culture of primary human OSE with endometrial stromal cells in an estrogen-rich environment.
- Gene expression analysis and functional assays (steroid conversion, SF-1 transfection) in immortalized cell lines.
- Tumorigenicity assessment of immortalized OSE in nude mice.
Main Results:
- Primary and immortalized OSE cells exhibited glandular-stromal structures resembling endometriosis and converted estrone to estradiol.
- OSE cells expressed genes for SF-1, p450arom, and 17beta-HSDs, indicating steroidogenic potential.
- One immortalized OSE clone formed tumors in nude mice, mimicking undifferentiated carcinomas.
- Immortalized granulosa cells (GC1a) required SF-1 transfection to achieve steroid hormone biosynthesis and estradiol secretion.
- SF-1 transgene induced 17beta-HSD enzymatic activity in GC1a cells, confirming its role in steroidogenesis.
Conclusions:
- Immortalized human OSE and granulosa cell lines provide valuable tools for studying ovarian physiology and pathology.
- OSE cells possess characteristics potentially linking them to endometriosis and exhibit estrogen metabolism capabilities.
- SF-1 is a key regulator of steroidogenic gene expression in human granulosa cells.
- Immortalized granulosa cells with steroidogenic capacity may be useful for in vitro oocyte maturation.
- Further research is needed to determine if OSE and granulosa cells share a common progenitor cell.
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