Gagliardi C L, Goldsmith L T, Saketos M, Weiss G, Schmidt C L
Department of Obstetrics and Gynecology, University of Medicine and Dentistry of New Jersey, Newark 07103.
Fertil Steril. 1992 Aug;58(2):314-20.
To determine the effect of human chorionic gonadotropin (hCG) on relaxin secretion by long-term cultures of luteinized human granulosa cells (GC).
Luteinized human GC were collected from 10 women undergoing in vitro fertilization (IVF) cycles. Luteinized human GC from each woman were plated in replicate wells at 1 x 10(5) cells/well and exposed to medium 199 (GIBCO, Grand Island, NY), medium 199 with 1 IU/mL hCG, and/or medium 199 with 100 IU hCG/mL. Luteinized human GC were maintained for up to 40 days in culture. Spent media were changed every 2 days and assayed for relaxin and progesterone (P) at the conclusion of each experiment.
Tertiary care center.
PATIENTS, PARTICIPANTS: Luteinized human GC were obtained from women undergoing controlled ovarian hyperstimulation for IVF with one of the following regimens: (1) clomiphene citrate with human menopausal gonadotropins (hMG); (2) hMG alone; or (3) hMG with leuprolide acetate. All women were less than 40 years of age, in good health, and were not taking medications other than those used in the ovulation-induction regimen.
Levels of P and relaxin in spent media.
Relaxin secretion by luteinized human GC was dependent on hCG stimulation and was detected only after a time lag in culture. After relaxin secretion was detected, it was maintained throughout the culture period (10 to 22 days). Luteinized human GC produced P immediately under both basal and stimulated conditions. Progesterone production continued throughout the culture period with hCG-stimulated cells producing significantly greater P after 4 to 8 days in culture.
Luteinized human GC obtained at the time of oocyte retrieval secrete relaxin in response to hCG stimulation and secrete P under both basal and hCG-stimulated conditions, thereby serving as a model to explore luteal function and control.
通过对人黄体化颗粒细胞(GC)进行长期培养,确定人绒毛膜促性腺激素(hCG)对松弛素分泌的影响。
从10名接受体外受精(IVF)周期的女性中收集黄体化人GC。将每名女性的黄体化人GC以1×10(5) 个细胞/孔的密度接种于多个复孔中,并分别暴露于199培养基(GIBCO,纽约州大岛)、含1 IU/mL hCG的199培养基和/或含100 IU hCG/mL的199培养基中。黄体化人GC在培养中维持长达40天。每2天更换一次培养液,并在每个实验结束时检测松弛素和孕酮(P)。
三级医疗中心。
患者、参与者:黄体化人GC取自接受IVF控制性卵巢过度刺激的女性,采用以下方案之一:(1)氯米芬与人绝经期促性腺激素(hMG)联合使用;(2)单独使用hMG;或(3)hMG与醋酸亮丙瑞林联合使用。所有女性年龄均小于40岁,身体健康,除排卵诱导方案中使用的药物外未服用其他药物。
培养液中P和松弛素的水平。
黄体化人GC分泌松弛素依赖于hCG刺激,且仅在培养一段时间后才能检测到。在检测到松弛素分泌后,其在整个培养期(10至22天)内持续存在。黄体化人GC在基础和刺激条件下均立即产生P。在整个培养期内,hCG刺激的细胞在培养4至8天后产生的P显著增加。
在卵母细胞采集时获得的黄体化人GC在hCG刺激下分泌松弛素,并在基础和hCG刺激条件下分泌P,从而作为探索黄体功能和调控的模型。