Follicle Stimulating Hormone Stimulation of 125l-Human Chorionic Gonadotropin Binding in Porcine Granulosa Cell Cultures 1
Follicle Stimulating Hormone Stimulation of 125l-Human Chorionic Gonadotropin Binding in Porcine Granulosa Cell Cultures 1
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猪颗粒细胞培养物中卵泡刺激激素刺激 125l-人绒毛膜促性腺激素结合 1
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发表时间：
1975
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通讯作者：
C. Channing
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作者：
C. Channing
为了观察FSH是否直接作用于颗粒细胞以刺激hCG结合，将从1-2 mm猪小卵泡收获的颗粒细胞在250 ml烧瓶中在含有0.05 μg/ml高度纯化的人FSH的化学成分确定的培养基中生长2、4和6天。确定的培养基由培养基199加0.4%牛血清白蛋白、0.2%乳清蛋白水解物和10 munit/ml胰岛素组成。用橡胶刮片刮取培养物，并与0.1 μg/ml 131 I-或125 I-hCG孵育。以cpm/培养物或每mg蛋白质表示的结合产生类似的结果。在五个独立的实验中，加入FSH刺激hCG结合，是对照培养物的2 - 4倍。在培养2天后的典型实验中，对照培养物与hCG的特异性结合为962 ± 45 cpm/培养物（x ± SE; n = 3），而在0.05 μg/ml FSH存在下生长的培养物中的结合为3933 ± 1787（n = 3; P < 0.01）。从在类似条件下生长的大（8-12 mm）卵泡收获的颗粒细胞结合率为29，669 ± 948 cpm/培养物（n = 4）。这些数据表明，FSH在体外诱导颗粒细胞LH-hCG受体后可能具有直接刺激作用。
Summary In order to see if FSH acts directly upon the granulosa cell to stimulate hCG binding, granulosa cells harvested from small 1-2 mm porcine follicles were grown in 250 ml flasks in chemically defined media containing 0.05 μg/ml highly purified human FSH for 2, 4, and 6 days. The defined medium consisted of culture medium 199 plus 0.4% bovine serum albumin, 0.2% lactalbumin hydrolysate and 10 munit/ml insulin. The cultures were harvested by scraping with a rubber policeman and incubated with 0.1 μg/ml 131I- or 125I-hCG. Binding expressed as cpm/culture or per mg protein yielded similar results. In five separate experiments addition of FSH stimulated hCG binding two- to fourfold above control cultures. In a typical experiment after 2 days of culture, the specific binding of control cultures to hCG was 962 ± 45 cpm/culture (x ± SE; n = 3) and the binding in cultures grown in the presence of 0.05 μg/ml FSH was 3933 ± 1787 (n = 3; P < 0.01). Granulosa cells harvested from large (8-12 mm) follicles grown under similar conditions bound 29,669 ± 948 cpm/culture (n = 4). These data demonstrate that FSH may have a direct stimulatory role upon induction of granulosa cell LH-hCG receptors in vitro.