培养的黄体化颗粒细胞释放的激素模拟了接受体外受精-胚胎移植的患者体内观察到的差异。

Hormone release from cultured luteinized-granulosa cells mimics differences seen in vivo in patients undergoing IVF-ET.

作者信息

Prien S D, Canez M S, Messer R H

机构信息

Department Ob/Gyn, TTUHSC, Texas Tech University Health Sciences Center, Lubbock 79430, USA.

出版信息

J Assist Reprod Genet. 1995 Mar;12(3):180-6. doi: 10.1007/BF02211795.

DOI:10.1007/BF02211795

PMID:8520182

Abstract

OBJECTIVES

Previous research from this laboratory has suggested that a relationship exists between the increase in circulating progesterone concentrations at the time of hCG administration and cycle outcome in patients undergoing IVF. Progesterone (P) increases of threefold or better within the 24-h period surrounding hCG administration appeared to be associated with a higher pregnancy rate. These data suggest a functional difference in the luteinized-granulosa of patients undergoing IVF. To test this hypothesis:

DESIGN

A split-split plot arrangement of treatments with two cell sources under five hormonal stimulations at four time points.

METHODS

Luteinized-granulosa cells (LGC) were collected from patients with either a normal increase (> or = threefold = NC) in circulating P (n = 4) or those with lower P increases (AC; n = 4). The cells were isolated by Ficoll gradient centrifugation and then cultured in 24-well culture plates using a modified media 199 containing 100 mIU/ml of hMG, FSH, LH, hCG, or a nonhormonal control to stimulate steroid-hormone production. At time points 2, 4, 6, and 8 days, media from each well was frozen for later hormone assay and replaced with fresh media containing the same stimulating factor. After all the media had been collected, P and estradiol (E2) released into the media were measured using radioimmunoassay.

RESULTS

Results indicate a higher media concentration of P (P < 0.001), but not E2 (P = 0.254), from NC, regardless of hormone stimulation or time in culture, when compared to the media from AC. Media concentrations of E2 were affected by a cell source by hormone stimulation by time interaction (P < 0.001) with varying effects. Media from NC maintained a constant E2 of between 1000-3000 pg/ml over the 8-day period (P = 0.163). However, media from AC demonstrated a stimuli-dependent E2 release (P < 0.001) ranging from < 1000 to over 11,000 pg/ml.

CONCLUSIONS

These data appear to support the existence of functionally different populations of luteinized-granulosa cells from patients undergoing IVF-ET.

摘要

目的

本实验室先前的研究表明，在接受体外受精（IVF）的患者中，注射人绒毛膜促性腺激素（hCG）时循环孕酮浓度的升高与周期结局之间存在关联。在hCG注射前后24小时内孕酮（P）浓度升高三倍或更高似乎与较高的妊娠率相关。这些数据表明接受IVF的患者的黄素化颗粒细胞存在功能差异。为验证这一假设：

设计

采用裂区裂区设计，在四个时间点对两种细胞来源进行五种激素刺激。

方法

从循环P正常升高（>或=三倍=NC；n = 4）的患者或P升高较低（AC；n = 4）的患者中收集黄素化颗粒细胞（LGC）。通过Ficoll梯度离心分离细胞，然后在24孔培养板中使用含有100 mIU/ml人绝经期促性腺激素（hMG）、促卵泡激素（FSH）、促黄体生成素（LH）、hCG或非激素对照的改良培养基199进行培养，以刺激类固醇激素的产生。在第2、4、6和8天，将每个孔中的培养基冷冻以备后续激素检测，并用含有相同刺激因子的新鲜培养基替换。收集所有培养基后，使用放射免疫分析法测量释放到培养基中的P和雌二醇（E2）。

结果

结果表明，与AC组的培养基相比，无论激素刺激或培养时间如何，NC组培养基中的P浓度更高（P < 0.001），但E2浓度无差异（P = 0.254）。E2的培养基浓度受到细胞来源、激素刺激和时间相互作用的影响（P < 0.001），且影响各异。NC组的培养基在8天内E2浓度维持在1000 - 3000 pg/ml之间（P = 0.163）。然而，AC组的培养基显示出刺激依赖性的E2释放（P < 0.001），范围从< 1000到超过11,000 pg/ml。