Effects of Ochratoxin A exposure on DNA damage in porcine granulosa cells in vitro

2020 ◽  
Vol 330 ◽  
pp. 167-175
Author(s):  
Tian-Yu Zhang ◽  
Li Kong ◽  
Jia-Xing Hao ◽  
Han Wang ◽  
Zi-Hui Yan ◽  
...  
1995 ◽  
Vol 83 (2-3) ◽  
pp. 169-177 ◽  
Author(s):  
Béatrice Goxe ◽  
Jacques E. Flechon ◽  
Solange Delasalle ◽  
Roland Salesse

1979 ◽  
Vol 80 (1) ◽  
pp. 9-20 ◽  
Author(s):  
ADA M. LINDSEY ◽  
CORNELIA P. CHANNING

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.


2012 ◽  
Vol 58 (4) ◽  
pp. 438-444 ◽  
Author(s):  
Malgorzata DUDA ◽  
Malgorzata DURLEJ ◽  
Malgorzata KNET ◽  
Katarzyna KNAPCZYK-STWORA ◽  
Zbigniew TABAROWSKI ◽  
...  

1988 ◽  
Vol 66 (5) ◽  
pp. 561-566 ◽  
Author(s):  
K. Rajkumar ◽  
P. Klingshorn ◽  
P. J. Chedrese ◽  
B. D. Murphy

Porcine granulosa cells cultured under serum free conditions responded by increased progesterone secretion to the addition of the leuteotropic hormones, LH, prolactin, and estradiol. Provision of extracellular substrate for steroidogenesis in the form of porcine high density lipoprotein or low density lipoprotein enhanced progesterone accumulation by granulosa cell cultures. Estradiol, LH, and prolactin all greatly increased progesterone accumulation in the presence of either high or low density lipoproteins. Increases in progesterone accumulation following addition of prolactin or LH in combination with estradiol suggested the presence of a synergistic interaction among leuteotropins. Pre-exposure of granulosa cell cultures to estradiol increased the subsequent stimulatory effect of prolactin on lipoprotein utilization. It is concluded that all three leuteotropins function to enhance and may interact in the utilization of extracellular lipoprotein substrate for progesterone synthesis.


2009 ◽  
Vol 46 (4) ◽  
Author(s):  
Małgorzata Durlej ◽  
Małgorzata Duda ◽  
Katarzyna Knapczyk ◽  
Maria Słomczyńska

2005 ◽  
Vol 18 (4) ◽  
pp. 677-681 ◽  
Author(s):  
E. Benzoni ◽  
M.L. Torre ◽  
M. Faustini ◽  
S. Stacchezzini ◽  
F. Cremonesi ◽  
...  

Three-dimensional culture systems in barium alginate capsules can be employed to maintain primary granulosa cells in an undifferentiated state for almost 6 days. This is due to a self-organization of cells in a pseudofollicular structure. The transfection of primary granulosa cells is a necessary condition when employing these culture systems for several purposes, for example as an in vitro toxicity test or the development of oocytes or zygotes. In this work, the feasibility of two transient transfection techniques (liposome-mediated and electroporation) was assessed in primary porcine granulosa cells after a 6-day culture in an artificial extracellular matrix (barium alginate membrane). Human recombinant green fluorescent protein was chosen as a molecular readout, and protein expression was assessed after 48 hours from transfection. Liposome-mediated transfection gave low transfection levels, with increasing yields from 2 to 12 μgDNA/ml of medium; the maximum percentage (85.7%) was reached at 12 μgDNA/ml of medium. Electroporation-mediated transfection yields were higher: the best results (81.7% of transfected cells) were achieved with two 50V pulses and 12 μg/ml DNA. The application of a single or double pulse (50V) at 4 mgDNA/ml gave negligible results. These results indicate that primary granulosa cell cultured in barium alginate capsules can be transfected by electroporation with high transfection yields.


2010 ◽  
Vol 22 (9) ◽  
pp. 134
Author(s):  
R. Smits ◽  
D. T. Armstrong ◽  
L. Ritter ◽  
M. Mitchell ◽  
M. B. Nottle

Caughey et al (2005) reported that prostaglandins derived from omega 3 sources eicsapentaenoic acid (EPA C20:5) and docosahexaenoic acid (DHA C22:6) have different properties to those derived from the preferred substrate, arachidonic acid (ARA C20:4; n-6). Armstrong et al (2006) demonstrated that PGE2 increased progesterone when porcine granulosa cells were cultured in vitro with hCG. We hypothesized that PGE3 which is derived from EPA will produce a lower steroidogenic response as progesterone from isolated granulosa cells collected from pre-ovulatory sow ovaries. Ovaries were collected from slaughtered sows and follicles between 3–8 mm were aspirated and through a series of wash steps in HTCM (Hepes TCM 199). Mid-sized granulosa cells were recovered in a solution of BTCM (bicarbonate TCM) containing IGF-1. 0.5 × 106 cells/mL were cultured in 250 µL of Control (BTCM only), PGE2 or PGE3 (320 ng/mL in BTCM, Cayman Chemical Co.) treatments with IGF1 at 25 ng/mL. Cultures were incubated for 22 h at 38oC. Cultures were centrifuged and the supernatant was analysed in duplicate for progesterone. Data was analysed by Univariate GLM ANOVA. There was no significant difference between PGE2 and PGE3 treatments, however the main effect of PGE significantly increased progesterone production relative to the control (P = 0.017). Granulosa cells cultured with omega 3 derived PGE3 did not produce significantly lower progesterone levels than those with PGE2. We conclude that both PGE2 and PGE3 promote a steroidogenic response in cultured porcine granulosa cells. (1) Armstrong DT, Formosa, ER, Amato F, Schultz SJ. 2006. Prostaglandin E2 up-regulates luteinizing hormone receptor (LHR) expression and enhances steroidogenic responses of follicle cells.(2) Caughey GE, James MJ, Cleland LG. 2005. Prostaglandins and leukotrienes. pp. 42–49. In ‘Encyclopaedia of Human Nutrition. Vol. 4’. (Eds B Caballero, L Allen, A Prentice).


1989 ◽  
Vol 120 (2) ◽  
pp. 251-260 ◽  
Author(s):  
M. W. Khalil ◽  
P. Morley ◽  
M. A. Glasier ◽  
D. T. Armstrong ◽  
T. Lang

ABSTRACT The origin and biosynthesis of 4-oestrene-3,17-dione (19-norandrostenedione), a major steroid in porcine ovarian follicular fluid, was investigated by culturing granulosa cells from 4–6 mm follicles of prepubertal gilts with radiolabelled androstenedione and 19-hydroxyandrostenedione. Steroid metabolites were purified by solvent extraction and lipophilic column chromatography, and analysed by C18 reverse-phase high-performance liquid chromatography. 19-Hydroxyandrostenedione, 19-norandrostenedione and oestradiol-17β were obtained as major metabolites from androstenedione, while 19-norandrostenedione and oestradiol-17β were the major products from 19-hydroxyandrostenedione. Serum alone or serum plus FSH significantly enhanced formation of 19-norandrostenedione and oestradiol-17β from each substrate, compared with controls. Micromolar concentrations (1 μmol/l) of 4-hydroxyandrostenedione, an aromatase inhibitor, significantly reduced formation of 19-norandrostenedione and oestradiol-17β by granulosa cells cultured with serum and FSH. Formation of 19-norandrostenedione and oestradiol-17β from androstenedione and 19-hydroxyandrostenedione was also significantly inhibited by aminoglutethimide phosphate, a cytochrome P-450 inhibitor known to block the conversion of androstenedione to oestrogens. Ketoconazole, an inhibitor of the cytochrome P-450 dependent 17,20-lysase, blocked formation of 19-norandrostenedione and oestradiol-17β only at millimolar concentrations. These results suggest that 19-norsteroid and oestrogen formation from C19 aromatizable androgens may share a common or overlapping pathway, and imply that 19-norsteroid and oestrogen synthesis is mediated by cytochrome P-450 dependent enzymes. Journal of Endocrinology (1989) 120, 251–260


1993 ◽  
Vol 5 (1) ◽  
pp. 95
Author(s):  
RT Denkova ◽  
IG Ivanov ◽  
LN Kanchev

The ability of porcine granulosa cells to release a progesterone inhibiting substance(s) was examined in vitro. Granulosa cells (SGCs, MGCs and LGCs) were harvested from small, medium or large antral follicles respectively. The effect of granulosa cell conditioned media obtained from small follicles (SGCCM) was studied in the culture of LGCs by estimation of progesterone secretion; the conditioned media evoked the inhibition of progesterone secretion by the LGCs. SGCCM produced by various numbers of cultured granulosa cells showed a dose-related inhibition of progesterone production. A maximum inhibitory effect was noted when a 5-fold concentration of SGCCM was used. The addition of SGCCM had no effect on the growth of the cultured cells. The factor(s) inhibiting progesterone secretion appeared to be a nonsteroidal substance of molecular mass greater than 10 kDa and was heat-stable and trypsin-sensitive. The data presented support the suggestion that the conditioned media generated by primary cultures of SGCs contain nonsteroidal regulators capable of inhibiting progesterone secretion by cultured LGCs; this inhibitory activity can play an important autocrine regulatory role in the process of follicular differentiation.


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