Granulosa cells culture

Steward, D. R., and Vande Voort, C. A., Relaxin secretion by human granulosa cell culture is predictive of in-vitro fertilization-embryo transfer success. Hum. Reprod. 14, 338-344 (1999). 

Haney, A.F., S.F. Hughes, and C.L. Hughes, Jr. 1984. Screening of potential reproductive toxicants by use of porcine granulosa cell cultures. Toxicology 30(3) 227- 241. 

Armstrong DT Dorrington JH (1976) Androgens augment FSH-induced progesterone secretion by cultured rat granulosa cells. Endocrinology, 99 1411. 

Lucky AW, Schreiber JR, Hillier SG, Schulman JD, 4 Ross GT (1977) Progesterone production by cultured preantral rat granulosa cells stimulation by androgens. Endocrinology, 100 128-133. 

LovekampTN, Davis BJ. 2001. Mono-(2-ethlhexyl) phthalate suppresses armotase transcript levels and estradial production in cultured rat granulosa cells. Toxicol Appl Pharmacol 172 217-224. 

FSH treatment in vitro also causes the formation of microvilli in granulosa cells and it has been suggested that LH receptors are localized mainly on microvilli. Accompanied by the FSH stimulation of steroidogenesis in cultured granulosa cells, morphological studies indicate the presence of smooth endoplasmic reticulum, mitochondria with tubular cristae, lipid droplets and Golgi apparatus in these cells [59],... 

Inhibin, an ovarian protein hormone, has been shown to selectively decrease FSH but not LH secretion by the anterior pituitary cell. The gonadal source of inhibin has been localized to the testis Sertoli cells [76] and ovarian granulosa [77]. In cultured granulosa cells, FSH increases inhibin production via stimulation of the protein kinase A pathway. In vivo treatment with injections of gonadotropin also increases the circulating levels of inhibin [78], whereas treatment with inhibin... 

In vitro, granulosa cells can be cultured and secrete hormones. Estradiol and progesterone secreted in vitro by granulosa cells are not correlated with treatment outcome, but relaxin secretion is correlated with conception and with term pregnancy (S25). Relaxin has a stimulating effect on endometrium, and this might be the reason for the reported correlation. 

Seifer, D. B., Gardiner, A. C., Lambert-Messerlian, G., and Schneyer, A. L., Differential secretion of dimeric inhibin in cultured luteinized granulosa cells as a function of ovarian reserve. J. Clin. Endocrinol. Metab. 81, 736-739 (1996). 

Culture of human cumulus granulosa cells (protocol n°92)... 

Studies in rodents have focused on the induction of ovarian NO synthesis primarily by cytokines. Rat ovarian NO production increases in response to hCG and IL-ip in the perfused ovary (Bonello et al., 1996) and in vivo (Nakamura et al., 1996). Cultured immature rat ovarian cells (Ben-Shlomo et al., 1994 Ellman et al., 1993) and isolated rat granulosa cells (Tabraue et al., 1997) synthesize NO in a dose-dependent manner in response to IL-ip. Furthermore, TNFa stimulates the NO pathway in isolated bovine thecal cells, causing increased cGMP levels, the significance of which is currently not clear (Tabraue et al., 1997). Taken together, these data demonstrate that the NO pathway is operative and sensitive to cytokine stimulation during the periovulatory period and suggests that NO is a downstream mediator of LH/hCG and IL-1 p in the ovulatory process. 

In cultured porcine granulosa cells, NO has been implicated as a negative regulator of steroidogenesis (Masuda et al., 1997). Treatment of purified granulosa cells with an NO donor resulted in decreased aromatase activity (Masuda et al., 1997). Furthermore, inhibition of NOS resulted in a dose-dependent increase in estradiol synthesis from both basal and gonadotropin-stimulated cells (Masuda et al., 1997). 

Dave, S Faramce, D.P., and Whitehead, S.A. (1997). Evidence that nitric oxide inhibits steroidogenesis in cultured rat granulosa cells. Clin. Sci. 92 277-284. 

Masuda, M., Kubota, T., Kamada, S., and Aso, T. (1997). Nitric oxide inhibits steroidogenesis in cultured porcine granulosa cells. Mol. Human Reprod. 5 285-292. 

Santana, P., Llanes, L., Hernandez, I., Gallardo, G., Quintana, J., Gonzalez, J., Estevez, F., Ruiz de Galarreta, C., and Fanjul, L. F. (1995). Ceramide mediates tumor necrosis factor effects on P450-aromatase activity in cultured granulosa cells. Endocrinology 136, 2345-2348. 

Treinen KA, Sneeden JL, Heindel JJ. 1993. Specific inhibition of cAMP accumulation by A -tetrahydrocannabinol in cultured rat granulosa cells. Toxicol Appl Pharmacol 118 53-57. 

Gonadotropin. Gonadotropin effects on PG secretion have not yet been studied extensively, but in our experiments, the exogenous luteinizing hormone (LH) was a potent stimulator of PGF release by cultured bovine granulosa cells (11 Fig. 1). 

Oxytocin, Oxytocin is a known stimulator of PGF secretion by the uterus. Moreover, OT-induced labor is due in part to a dramatic increase in the production of PGF, which stimulates uterine contractions (1). Oxytocin was shown to be an activator of PGF release by cultured porcine ovarian follicles (8), and porcine (Fig. 2) and bovine (10 Fig. 1) granulosa cells. Oxytocin also stimulated PGE secretion by porcine ovarian follicles (8 Fig. 3), hamster ovarian cells (9), and bovine granulosa cells (11 Fig. 1). The immunoneutralization of OT produced by porcine granulosa cells inhibited PGE release (10). Thus, OT has been shown to be a stimulator of ovarian PGF and PGE secretion. 

Fig. 2. Effect of oxytocin (0,1,10 or 100. 1.000 or 10.000 ng/mL medium) on prostaglandin (PG)F release by cultured porcine ovarian granulosa cells. Values are means SEM. Sig-nificant differences (P< 0.05) between cells cultured with and without exogenous oxytocin. Methods of isolation and culture of ovarian granulosa cells, as well as the PGF radioimmunoassay are described in (10).

Sirotkin, A.V., and Makarevich, A.V. (1999) GH Regulates Secretory Activity and Apoptosis in Cultured Bovine Granulosa Cells Through the Activation of the cAMP/Protein Kinase A System, J. Endocrinol. 163,317-327. 

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