Considering all these results, we propose the hypothesis that the appearance of theca cell layers and the blood vessels in them are key factors initiating the proliferation of granulosa cells in the early stage of follicular development. In the present study, bovine ovarian stromal cells, which had neither LHCGR mRNA nor hCG responsiveness, were used to examine the process by which stromal cells differentiate into steroidogenic cells and acquire the capacity to respond to gonadotropins. Using the immature hypophysectomized rat model, Huang et al. Moreover, our observations also suggest that granulosa-theca cell interactions are essential for the maintenance of normal follicular structure and function [9–11]. Whether the continued growth of the TC layer is caused by continued effects of GCs on additional cortical stromal cells and/or by simple proliferation of existing TCs remains to be determined. 3rd Ed. Briefly, granulosa cells were harvested by aseptic needle aspiration from follicles and washed three times with a culture medium consisting of Waymouth's MB 752/1 medium (Gibco, Grand Island, NY), Hanks' solution (Gibco), and 10% fetal calf serum (FCS) (6:3:1, v:v:v) supplemented with 100 μg/ml streptomycin and 100 U/ml penicillin (Gibco). Ovaries were placed in an ice-cold buffered salt solution and transferred to the laboratory less than 60 min after collection. Differences between treatment groups were analyzed using Tukey or Bonferroni post-test; significance was inferred for P < 0.05. This effect of the theca cells, however, was not observed in the present study. Gougeon A. Dynamics of human follicular growth: a morphologic perspective. To study the effect of GCs on ovarian stromal cells, isolated SC and SM were cultured with or without GCs in serum-supplemented medium for 48 h. The morphology of the cultured stromal cells was examined using an electron microscope. Nonsteroidal signals originating in the gonads. The tumors without duplications carried point mutations affecting highly conserved residues. The other possibility is that theca cells send the same factor(s) to granulosa cells throughout the follicular maturation process and that granulosa cells from small- and large-sized follicles show different types of responses to the same theca factor(s). Plating density(number of cells per volume of culture medium) plays a critical role for the differentiation. The granulosa lutein cells do have aromatase, and use it to produce estrogens, using the androgens previously synthesized by the theca lutein cells, as the granulosa lutein cells in themselves do not have the 17α-hydroxylase or 17,20 lyase to produce androgens. Regulation of ovarian cell growth through the local production of transforming growth factor-α by theca cells. The second antibody for the inhibin assay was kindly provided by Dr. Katsumi Wakabayashi (Biosignal Research Center, Institute for Molecular and Cellular Regulation, Gumma University). The theca factor(s) inhibits the differentiation of granulosa cells at the early stage of follicular development but promotes it during late follicular maturation, while the granulosa factor(s) promotes both differentiation and growth of theca cells throughout the follicular maturation process. Cholesterol is released and stored within the cell as cholesterol ester. It has generally been accepted that growth of the ovarian follicle is under the endocrine control of the pituitary gonadotropins. Plating density (number of cells per volume of culture medium) plays a critical role for the differentiation. For this study, ovaries with a corpus luteum light yellow to white in color, < 1 cm in diameter, and avascular surface were used. Androstenedione production by 1 × 105 theca cells cultured alone (T-alone) and with granulosa cells (T/G) during the first and the second 48 h of culture in a serum-free medium. Effect of GCs on hCG responsiveness of cultured ovarian stromal cells. Ovarian stromal cell differentiation is characterized by increased gene expression for androgenic factors [26]. These observations support the notion that the ovarian stroma is the cellular origin of TCs and that GCs stimulate this differentiation process. Messenger RNA expression for CYP17A1 (30 cycles), CYP11A1 (30 cycles), HSD3B1 (30 cycles), LHCGR (30 cycles), STAR (30 cycles), NR5A1 (30 cycles), VEGF (30 cycles), and ARBP (internal control; 25 cycles) were examined using RT-PCR assays. Total cellular RNAs were extracted with TRIzol (Invitrogen Corp.) by the guanidium acid-isothiocyanate-phenol-chloroform method, in accordance with the manufacturer's protocol. [5] It also produces relaxin, a hormone responsible for softening of the pubic symphysis which helps in parturition. In the presence of GCs, stromal cells also exhibited increased expression of mRNA for LHCGR and LH responsiveness, two well-established differentiated functions of TCs in the bovine ovary [27]. The uterine lining (endometrium) is expelled through the vagina (in mammals that go through a menstrual cycle). Our preliminary study showed that single-cultured theca cells from both small and large follicles produced negligible amounts of progesterone (the progesterone-producing ability of theca cells from small and large follicles was less than 5% and 6%, respectively, of that of single-cultured granulosa cells) and did not produce estradiol or inhibin. GCs and TCs were collected from small antral follicles (2–4 mm diameter), as described previously [19]. The ovarian disease has two forms, juvenile and adult, both characterized by indolent growth,[1] and therefore has high recovery rates. Decreased granular cell proliferation (growth) and increased cell death are described in granular cells obtained from infertile women with decreased ovarian reserve due to relationship between increasing FSH levels (suggestive of declining ovarian reserve) and declining granular cell viability. A P value of less than 0.05 was considered significant. Hence both the numbers and maturation of granulosa cells in any given follicle are important and both processes are regulated by gonadotrophic hormones from the anterior pituitary. [23], with slight modifications.
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