1996
DOI: 10.1262/jrd.42.133
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In Vitro Differentiation of Bovine Granulosa Cells Obtained from Small Antral Follicles.

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Cited by 4 publications
(5 citation statements)
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“…In the present study, we used a model of luteinizing granulosa cells induced by insulin (2 µg/ml) and forskolin (10 µM). Bovine granulosa cells obtained from small antral follicles are known to differentiate into large luteal-like cells during culture in vitro [ 25 ]. Our results showed that the cultured granulosa cells treated with insulin in combination with forskolin showed increased P4 production after 24 h, and the production was the highest among the groups ( Fig.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…In the present study, we used a model of luteinizing granulosa cells induced by insulin (2 µg/ml) and forskolin (10 µM). Bovine granulosa cells obtained from small antral follicles are known to differentiate into large luteal-like cells during culture in vitro [ 25 ]. Our results showed that the cultured granulosa cells treated with insulin in combination with forskolin showed increased P4 production after 24 h, and the production was the highest among the groups ( Fig.…”
Section: Discussionmentioning
confidence: 99%
“…To prepare luteinizing and non-luteinizing granulosa cells, the culture medium was replaced with fresh medium containing 0.1% BSA, 5 ng/ml sodium selenite (Sigma-Aldrich; S5261), 5 µg/ml transferrin (Sigma-Aldrich; T4132) and 0.5 mM ascorbic acid (Wako-Pure Chemical Industries, Osaka, Japan; 031–12061), and the cells were then incubated under a normal culture atmosphere (20% O 2 , 5% CO 2 , 75% N 2 ) without or with insulin (2 µg/ml; Sigma-Aldrich; I4011), forskolin (10 µM; Research Biochemicals International, Natick, MA, USA; 70–0501-05) or insulin (2 µg/ml) in combination with forskolin (10 µM) for 24 h. The concentration of insulin and forskolin was selected based on a previous report [ 25 ]. The culture media from these cultured cells were collected to determine the effect of insulin and forskolin treatment on P4 production for 24 h.…”
Section: Methodsmentioning
confidence: 99%
“…Insulin in combination with forskolin mimics the effects of luteinizing hormone (LH) and activates adenylate cyclase via upregulation of P4 [ 33 ]. The concentration of insulin and forskolin was selected according to other reports [ 26 , 34 ].…”
Section: Methodsmentioning
confidence: 99%
“…The P 4 concentration was determined d i r e c t l y i n t h e m e d i u m w i t h a n e n z y m e immunoassay modified [26] from the original method described by Prakash et al [26]. Briefly, progesterone 3-(O-carboxymethyl) oxime linked to peroxidase type VI through a 6-aminocaproic acid spacer (P3659, Sigma) was used as a tracer (1:120, 000 final dilution).…”
Section: Determination Of P 4 Concentration In Maturation Medium In Wmentioning
confidence: 99%