Molecular and Cellular Endocrinology 228 (2004) 67–78
Ovarian granulosa cell lines
Jon C. Havelock, William E. Rainey, Bruce R. Carr∗
Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology, University of Texas Southwestern Medical Center,
5323 Harry Hines Boulevard, Dallas, TX 75390-9032, USA
Received 21 January 2004; accepted 14 April 2004
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
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.
© 2004 Elsevier Ireland Ltd. All rights reserved.
Keywords: Granulosa cell; Folliculogenesis; Estrogen
The ovaries are complex endocrine organs developed
from the bipotential gonad (Gillman, 1948). In adult life
and under the stimulatory actions of the gonadotropins, they
are responsible for the production of the sex steroids and
are the source of fertilizable ova. The sex steroids act in an
autocrine/paracrine fashion in the human ovary to produce
the complex interactions necessary for folliculogenesis and
act in an endocrine fashion to regulate the orderly hormonal
ble for the secondary sexual characteristics that appear with
reproductive age. Reproductive disorders such as polycystic
ovarian syndrome (PCOS), manifested by chronic anovula-
tion and hyperandrogenism, are common in reproductive-
∗Corresponding author. Tel.: +1 214 648 4747; fax: +1 214 648 8066.
E-mail address: email@example.com (B.R. Carr).
disturbances. Greater understanding of the mechanisms that
regulate and control ovarian function and the perturbations
in ovarian steroidogenesis in these disorders may lead to
novel therapies for a variety of reproductive disorders.
The ovary is composed of three distinct regions: an outer
cortex containing the germinal epithelium and the follicles,
a central medulla consisting of stroma, and a hilum around
the area of attachment of the ovary to the mesovarium (Carr,
1998). The steroidogenic cells of the ovary are the granulosa
cells, which are the avascular cellular compartment sur-
rounding the oocyte, and the theca cells, which reside in the
ovarian stroma. These two cellular compartments are sepa-
rated by the basal lamina (Weakly, 1966). The ovary secretes
a number of steroids, including pregnenolone, progesterone,
17?-hydroxyprogesterone, 17?-hydroxypregnenolone, de-
hydroepiandrosterone, androstenedione, testosterone, es-
trone, and 17?-estradiol (Baird et al., 1974; Baird and
Fraser, 1969). Under the control of various hormones,
second-messenger signalling such as cyclic AMP (cAMP),
cytokines and transcription factors, ovarian steroid produc-
0303-7207/$ – see front matter © 2004 Elsevier Ireland Ltd. All rights reserved.
J.C. Havelock et al. / Molecular and Cellular Endocrinology 228 (2004) 67–78
Fig. 1. Follicular phase steroid biosynthesis in the ovary with the illustra-
tion of the two-cell/two-gonadotropin theory. It should be noted that there is
some species variation in steroidogenic pathways within the ovary and the
clature for the enzymes: StAR, steroidogenic acute regulatory protein;
hydroxysteroid dehydrogenase type I; CYP19, aromatase.
tion qualitatively and quantitatively changes throughout the
ovulatory cycle, with increased steroidogenic activity in the
luteal phase, accompanied by a shift from a predominant
estrogen producing to a progesterone producing organ.
In humans, ovarian steroidogenesis occurs according
to the two-cell/two-gonadotropin theory (Ryan and Petro,
1966). For estrogen biosynthesis to occur, synthesis of
C19 androgens from cholesterol occurs in the luteinizing
hormone (LH) stimulated theca cell compartment, as CYP17
activity is predominantly limited to the theca cells (Sasano
et al., 1989) (Fig. 1). These androgens then diffuse into the
avascular, CYP17 deficient granulosa cell compartment.
Under follicle stimulating hormone (FSH) stimulation,
these androgens either undergo aromatization to estrogens
via aromatase (CYP19) activity (Bjersing, 1968), or are
preferentially 5?-reduced when present in an androgen-rich
environment (McNatty et al., 1979). In the luteal phase,
the luteinized granulosa cell compartment becomes vascu-
larized, allowing delivery of cholesterol to the previously
avascular granulosa cell compartment. Increased expression
of steroidogenic enzymes also occurs, resulting in increased
production of progesterone as well as estrogen derived from
theca cell androgens (Fig. 2). While this model also holds
for primates and some animals, variations exist, such as in
the porcine ovary, that demonstrate cellular differences in
steroidogenic enzyme expression, and as a result, steroido-
genic capability (Conley et al., 1995). In addition, many
Fig. 2. Steroidogenic pathways for luteinized theca and granulosa cells.
AC, adenylate cyclase; LH, luteinizing hormone. It should be noted that
there is some species variation in steroidogenic pathways within the ovary
and the pathway provided in this figure is characteristic of human. Some
species lose CYP17 expression in theca-lutein cells, resulting in proges-
terone synthesis. AC, adenylate cyclase; FSH, follicle stimulating hormone;
LH, luteinizing hormone. Nomenclature for the enzymes: StAR, steroido-
genic acute regulatory protein; CYP11A, cholesterol side-chain cleavage;
CYP17, 17?-hydroxylase, 17,20-lyase; HSD3B2, 30-hydroxysteroid dehy-
drogenase type II; HSD17B1, 17?-hydroxysteroid dehydrogenase type I;
animal species lose the ability to produce estrogen in the
luteal phase. Thus, ovarian steroid production has evolved a
number of species-specific differences in cell function.
Understanding the mechanisms which regulate ovarian
lular level requires readily available cells for in vitro stud-
ies. For humans, the major source of granulosa cells for in
(IVF). These cells are limited in number, and under stimula-
tion with supraphysiologic doses of FSH and human chori-
onic gonadotropin (hCG) in vivo, they have a limited life
perstimulation with gonadotropins, these cells are normally
fullly luteinized at the time of isolation. Because of this, uti-
lizing primary cultures of human granulosa cells for research
ficult to obtain. The use of theca cell models through long-
term theca-cell cultures or tumor cell models has allowed for
greater understanding of these androgen producing cells, but
development of thecal cell lines has met with limited success
in vitro model systems has become an attractive alternative.
J.C. Havelock et al. / Molecular and Cellular Endocrinology 228 (2004) 67–78
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