Effects of growth hormone on FSH, insulin and triiodothyronine-mediated estradiol production by granulosa cells from prepubertal gilts in vitro

1993 ◽  
Vol 73 (2) ◽  
pp. 443-447 ◽  
Author(s):  
K. Rajkumar ◽  
R. N. Kirkwood ◽  
P. A. Thacker
Keyword(s):  
Growth Hormone ◽  
Granulosa Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Viable Cells ◽  
Porcine Granulosa Cells ◽  
The Media ◽  
Dose Dependent ◽  
Production Response

Three experiments were performed using porcine granulosa cells from medium-sized follicles (2–4 mm) cultured in serum-free medium at a density of 2 × 106 viable cells per well. Cells were cultured in the presence of combinations of FSH, insulin, growth hormone (GH) and triiodothyronine (T3), and subsequent estradiol (E2) production was monitored. In the presence of insulin, a dose-dependent E2 production response to FSH was observed (exp. 1). However, in all experiments, E2 production was reduced (P < 0.05) by the inclusion of GH in the media. The inclusion of T3 augmented the E2 response to insulin, an effect attenuated by GH. We conclude that GH, at the dose employed, is inhibitory to E2 production in granulosa cells of medium-sized ovarian follicles. Key words: Granulosa cells, growth hormone, estradiol

Effect of noradrenaline and dopamine on progesterone and estradiol secretion of human granulosa cells

1993 ◽  
Vol 129 (2) ◽  
pp. 165-168 ◽  
Author(s):  
József Bódis ◽  
Hans R Tinneberg ◽  
Attila Török ◽  
Philippe Cledon ◽  
Volker Hanf ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Direct Action ◽  
Serum Free Medium ◽  
Free Medium ◽  
Vitro Fertilization ◽  
Serum Free ◽  
Human Granulosa Cells ◽  
Estradiol Secretion ◽  
Cells Cultured

The aim of this study was to explore the direct action of noradrenaline and dopamine on progesterone and estradiol secretion of human granulosa cells cultured in serum-free medium. Progesterone and estradiol production was measured in the presence and absence of noradrenaline, dopamine or propranolol using radioimmunoassays; statistical analysis was performed by analysis of variance and Newman-Keul's multiple range test. Twenty-six women aged 31±3 years undergoing in vitro fertilization and embryo transfer for infertility treatment at University Women's Hospital, University of Tübingen, Germany, took part in this study. Noradrenaline significantly inhibited progesterone production by human granulosa cells in a dose-related manner at a concentration of 10−4–10−6 mol/l. Dopamine significantly stimulated estradiol secretion by granulosa cells in an inverse dose-related manner. Both effects were blocked by propranolol. The results suggest that catecholaminergic actions switch over the steroid production of human granulosa cells cultured in serum-free medium from progesterone to estradiol.

Effects of growth hormone and triiodothyronine on insulin-induced progesterone production by granulosa cells from prepubertal gilts

1992 ◽  
Vol 72 (3) ◽  
pp. 589-593 ◽  
Author(s):  
R. N. Kirkwood ◽  
P. A. Thacker ◽  
K. Rajkumar
Keyword(s):  
Growth Hormone ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Bovine Insulin ◽  
Dependent Manner ◽  
Progesterone Production ◽  
Porcine Granulosa Cells

Two experiments were performed using granulosa cells from medium-sized follicles (2–4 mm) derived from prepubertal gilts. Cells were cultured in a serum-free medium at a density of either 1 or 2 × 106 viable cells per well (experiments 1 and 2, respectively). For exp. 1, porcine growth hormone (pGH) (0 or 100 ng mL−1) was included in the culture medium from the time of plating, and low-density lipoprotein (LDL) (100 μg mL−1) was added at 72 h. For exp. 2, granulosa cells were plated in a culture medium containing either pGH (0 or 100 ng mL−1) or triiodothyronine (T3) (0 or 5 ng mL−1) or both pGH T3; LDL was not included. For both experiments, after 24 h of culture, bovine insulin at 0, 10, 100 or 1000 ng mL−1 was included in the medium. Hormones were replaced at 48 and 72 h, and the cultures were terminated at 96 h. Results from exp. 1 indicated that insulin increased (P < 0.01) progesterone production in a dose-dependent manner, both in the presence and absence of LDL. This response was augmented (P < 0.01) by co-culture with pGH. Results from exp. 2 confirmed the augmenting effect of pGH (P < 0.01). It was further observed that T3 increased (P < 0.01) progesterone production when cultured with insulin at 1000 ng mL−1, but at lower insulin-inclusion levels, results were equivocal. The progesterone production response was greatest (P < 0.01) when cells were cultured with both pGH and T3 at insulin levels of 100 or 1000 ng mL−1. There appeared to be little relationship between the media concentrations of insulin-like growth factor 1 and progesterone. The present results suggest that relatively high levels of pGH and T3 will enhance the in vitro steroidogenic capabilities of porcine granulosa cells. Key words: Granulosa cells, GH, T3, insulin

Dose response of luteinized porcine granulosa cells in vitro to prolactin: dependency on pre-exposure to human chorionic gonadotrophin

1988 ◽  
Vol 66 (10) ◽  
pp. 1337-1340 ◽  
Author(s):  
Pedro J. Chedrese ◽  
Kadaba Rajkumar ◽  
Hoa Ly ◽  
Bruce D. Murphy
Keyword(s):  
Granulosa Cells ◽  
Calf Serum ◽  
Density Lipoprotein ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Cells In Culture ◽  
The Media ◽  
Free Media ◽  
Dose Dependent

This study examined the hypothesis that human chorionic gonadotrophin (hCG) increases prolactin (PRL) stimulation of the utilization of lipoprotein-borne cholesterol by pig luteinized granulosa cells in culture. These cells, which luteinize in culture, were harvested from 6-mm or greater diameter follicles and cultured in the presence of 1% fetal calf serum and 1 μ/mL insulin for 48 h. On the third day, the media were replaced with fresh serum-free media, with the same dose of insulin, and on the following day (day 4) the media were replaced with serum- and insulin-free media. At this time (day 4) hCG was added to some cultures. On day 5, cells from the group with hCG and cells from the group without hCG were treated with graded doses of ovine PRL (0.1–3.0 μg/mL). To a second set of cells, likewise treated, 100 μg of porcine low density lipoprotein (LDL) was added. Two days later (day 7) media were sampled and replaced with media alone or media containing hormones and (or) LDL. On day 9 cultures were terminated. In the cells pre-exposed to hCG, PRL (1 μg/mL) in the presence of LDL increased progesterone production 1.7-fold (p < 0.01) on day 7 and 2.2-fold (p < 0.01) on day 9. In the granulosa cells in culture pre-exposed to hCG, the effect of PRL on LDL utilization was dose dependent and saturable at 1 μg/mL on days 7 and 9. We conclude that brief pretreatment of luteinized pig granulosa cells with hCG results in a dose-dependent PRL-induced utilization of LDL for progesterone synthesis.

RGD-mediated adhesion of porcine granulosa cells modulates their differentiation response to FSH in vitro

1995 ◽  
Vol 83 (2-3) ◽  
pp. 169-177 ◽  
Author(s):  
Béatrice Goxe ◽  
Jacques E. Flechon ◽  
Solange Delasalle ◽  
Roland Salesse
Keyword(s):  
Granulosa Cells ◽  
Porcine Granulosa Cells

scholarly journals Quantification of Internalized Silica Nanoparticles via STED Microscopy

2015 ◽  
Vol 2015 ◽  
pp. 1-16 ◽  
Author(s):  
Henrike Peuschel ◽  
Thomas Ruckelshausen ◽  
Christian Cavelius ◽  
Annette Kraegeloh
Keyword(s):  
Silica Nanoparticles ◽  
Super Resolution ◽  
Engineered Nanoparticles ◽  
Stimulated Emission ◽  
Alveolar Type ◽  
Serum Free Medium ◽  
Free Medium ◽  
Essential Information ◽  
Serum Free

The development of safe engineered nanoparticles (NPs) requires a detailed understanding of their interaction mechanisms on a cellular level. Therefore, quantification of NP internalization is crucial to predict the potential impact of intracellular NP doses, providing essential information for risk assessment as well as for drug delivery applications. In this study, the internalization of 25 nm and 85 nm silica nanoparticles (SNPs) in alveolar type II cells (A549) was quantified by application of super-resolution STED (stimulated emission depletion) microscopy. Cells were exposed to equal particle number concentrations (9.2×1010particles mL−1) of each particle size and the sedimentation of particles during exposure was taken into account. Microscopy images revealed that particles of both sizes entered the cells after 5 h incubation in serum supplemented and serum-free medium. According to thein vitrosedimentation, diffusion, and dosimetry (ISDD) model 20–27% of the particles sedimented. In comparison, 102-103NPs per cell were detected intracellularly serum-containing medium. Furthermore, in the presence of serum, no cytotoxicity was induced by the SNPs. In serum-free medium, large agglomerates of both particle sizes covered the cells whereas only high concentrations (≥ 3.8 × 1012particles mL−1) of the smaller particles induced cytotoxicity.

COMPARISON OF THE STIMULATORY EFFECTS OF OVINE, PORCINE AND HUMAN FOLLICLE-STIMULATING HORMONE AND OF OVINE AND HUMAN LUTEINIZING HORMONE ON THE ACCUMULATION OF CYCLIC AMP BY PORCINE GRANULOSA CELLS

1979 ◽  
Vol 80 (1) ◽  
pp. 9-20 ◽  
Author(s):  
ADA M. LINDSEY ◽  
CORNELIA P. CHANNING
Keyword(s):  
Cyclic Amp ◽  
Granulosa Cells ◽  
Incubation Medium ◽  
Follicle Stimulating Hormone ◽  
Similar Response ◽  
Intracellular Accumulation ◽  
Porcine Granulosa Cells ◽  
Tenfold Increase ◽  
Incubation Periods

The effects of ovine, porcine and human FSH, and ovine and human LH on the accumulation of cyclic AMP by porcine granulosa cells obtained from follicles at various stages of maturation were investigated. During incubation periods of 15 min, 10 μg ovine FSH pretreated with antiserum to LH or 10 μg human FSH resulted in an 11- to 18-fold, five-to ninefold, and less than a twofold increase in intracellular accumulation of cyclic AMP by granulosa cells from small (1–2 mm), medium (3–5 mm) and large (6–12 mm) follicles respectively. Similar patterns of response occurred with addition of porcine FSH. After incubation for 30 and 60 min with ovine, porcine or human FSH, significant accumulation of cyclic AMP in the incubation medium occurred with cells obtained from small and medium-sized follicles. After 60 min of incubation with FSH the accumulation of cyclic AMP in the incubation medium exceeded the intracellular cyclic AMP levels in granulosa cells from small and medium-sized follicles. During incubation periods of 15 min, 1·0 μg ovine LH resulted in less than a twofold, a fourfold and greater than a tenfold increase in intracellular accumulation of cyclic AMP by granulosa cells from small, medium and large follicles respectively. Addition of human LH brought about a similar response. Incubation periods of 30 and 60 min with 1·0 μg ovine or human LH resulted in significant accumulation of cyclic AMP in the incubation medium by granulosa cells from large follicles; cyclic AMP content in the incubation medium was greater after 60 min compared with 30 min of incubation. It was concluded that ovine FSH pretreated with an antiserum to LH had similar effects on cyclic AMP levels as did purified human and porcine FSH, and that the stimulatory effects of the less pure ovine FSH were probably not due to an impurity in the FSH preparation. Porcine granulosa cells obtained from small follicles should be suitable as an in-vitro FSH bioassay while granulosa cells obtained from large follicles should be suitable as an in-vitro LH bioassay.

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