Article

Ovarian granulosa cell lines

Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9032, USA.
Molecular and Cellular Endocrinology (Impact Factor: 4.24). 01/2005; 228(1-2):67-78. DOI: 10.1016/j.mce.2004.04.018
Source: PubMed

ABSTRACT The ovary is a complex endocrine gland responsible for production of sex steroids and is the source of fertilizable ova for reproduction. It also produces various growth factors, transcription factors and cytokines that assist in the complex signaling pathways of folliculogenesis. The ovary possesses two primary steroidogenic cell types. The theca cells (and to a lesser extent, the stroma) are responsible for androgen synthesis, and the granulosa cells are responsible for conversion of androgens to estrogens, as well as progesterone synthesis. These cells undergo a transformation in the luteal phase of the menstrual cycle, converting them from estrogen producing, to predominantly progesterone producing cells. Understanding the molecular mechanisms regulating these cells is essential in understanding the regulation of steroidogenesis and reproduction. Creation of appropriate in vitro cell model systems can provide important tools for the study of ovarian function. This has led to the development of ovarian steroidogenic cell lines in several laboratories. Developing theca cell lines has met with limited success. Conversely, numerous human and animal granulosa cell lines have been developed. This review will discuss the existing granulosa cell lines and their characteristics.

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    • "Although cell lines are technically easier to work with when compared with primary culture, the former may acquire different gene expression patterns and no longer represent the characteristics of the physiological cellular states, particularly when they are manipulated by a range of methods including co-or triple transfection either with oncogenes or tumor suppressor genes (Havelock et al., 2004). Moreover, the validation of the granulosa cell culture as a representative of the different developmental stage of the follicle was not established, and therefore, its use as a model for ovarian studies is limited (Havelock et al., 2004). "
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    ABSTRACT: 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 4 days of incubation of luteinized-MGCs, ovulatory genes mRNA's achieve expression levels similar to the early non-luteinized follicles. FSH stimulation for 48 hours of these 4 days 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.
    Molecular and Cellular Endocrinology 02/2014; 384(1). DOI:10.1016/j.mce.2014.01.018 · 4.24 Impact Factor
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    • "Small molecular agonists of aromatase would be useful to promote local estrogen biosynthesis for the prevention of estrogen deficiency-induced diseases (osteoporosis, neurodegenerative diseases, and cardiovascular diseases), but have been rarely reported [12]. Therefore, we examined the effects of newly isolated compounds 1–11 on estrogen biosynthesis in human ovarian granulosa-like KGN cells [13] [14]. Due to the limited amounts of these compounds isolated, only the activity of compounds 1, 5, 7, 9 and 10 were tested and described herein. "
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    ABSTRACT: Nine new highly oxygenated stigmastane-type steroids, vernoanthelcin A-I (1-9), and two new stigmastane-type steroidal glycosides, vernoantheloside A and B (10 and 11) were isolated from the aerial parts of Vernonia anthelmintica Willd. The structures of compounds 1-11 were determined on the basis of IR, MS, 1D-NMR, and 2D-NMR, and their absolute configurations were deduced using single-crystal X-ray diffraction and the CD exciton chirality method. Compounds 1, 5, 7, 9 and 10 were tested for their effects on estrogen biosynthesis in human ovarian granulosa-like cells (KGN cells).
    Steroids 03/2012; 77(7):811-8. DOI:10.1016/j.steroids.2012.03.003 · 2.72 Impact Factor
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    • "Granulosa cells are described to produce estradiol when incubated with FSH in the presence of the steroid precursor 4-an- drostene-3,17-dione (AD) according to the two-cells/two-gonatropin theory (Havelock et al., 2004). KGN was confirmed to synthesize estradiol when incubated with FSH under AD supplementation . "
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    ABSTRACT: Follicle stimulating hormone (FSH) plays a central role for in vitro fertilization therapy. Historically, FSH was extracted from urine of post-menopausal women. Now, most preparations used in Europe and North America are of recombinant origin. For the past several decades, the potency of FSH dosage has been determined by a single in vivo potency assay based on weight gain of rat ovaries upon hormonal treatment. This assay lacks precision, is cumbersome in interpretation, and sacrifices large numbers of rats. We herewith present the development and qualification of an alternative potency assay that will circumvent the aforementioned obstacles. The setup is based on a human ovary granulosa cell lines expressing the intact human FSH receptor on its surface. FSH binding results in induction of progesterone expression and secretion, which is subsequently quantitatively determined by a commercially available diagnostic ELISA. We demonstrate in this publication that the new assay features excellent precision and accuracy and has a wider working range than the in vivo assay. Analyses of commercial products of different origin using the in vitro assay confirm their stated potency. Further, it was demonstrated that the assay delivered comparable results to the Steelman/Pohley assay for a newly developed recombinant FSH product. The assay has been successfully validated for that product. Its accuracy, handling, and universal application for FSH preparations of different origin qualify the presented assay to complement and replace the Steelman/Pohley assay in the future.
    Altex Proceedings, 1/12, Proceedings of WC8, Montreal, Canada; 08/2011
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