Mikkelsen A L, Høst E, Lindenberg S
The Fertility Clinic, Institute for Human Reproduction, Symbion, Fruebjergvej 3, DK-2100 Copenhagen, Denmark.
Reproduction. 2001 Sep;122(3):481-6. doi: 10.1530/rep.0.1220481.
The aim of this study was to investigate the incidence of apoptosis in granulosa cells from immature human follicles undergoing in vitro maturation (IVM) and to compare the incidence of apoptotic granulosa cells (i) between FSH-primed and unprimed normal ovaries and (ii) between polycystic and normal ovaries. Furthermore, the incidence of apoptosis was related to maturation and subsequent fertilization and cleavage of the oocytes from the corresponding ovary. Seventy women undergoing 70 IVM cycles were included. Group 1 consisted of patients with normal ovaries (n = 52) and group 2 consisted of patients with polycystic ovaries (n = 18). Patients in group 1 were subdivided into two groups according to priming with FSH before aspiration. In group 1a (n = 27 cycles) oocytes were obtained in unstimulated cycles. In group 1b (n = 25 cycles) oocytes were obtained after priming with recombinant FSH for 3 days initiated on day 3 after spontaneous menstruation. In group 2 all patients were primed with recombinant FSH for 3 days before aspiration. Aspiration was performed transvaginally and cumulus-enclosed oocytes were matured for 28-30 h before fertilization. Granulosa cells were collected from follicular aspirates. An APOPTAG detection kit was used to stain the granulosa cells and to detect apoptosis. The incidence of apoptosis in granulosa cells was decreased in follicles from FSH-primed normal ovaries compared with follicles from unprimed normal ovaries and FSH-primed polycystic ovaries. No difference was found between granulosa cells from FSH-primed polycystic ovaries and granulosa cells from unstimulated normal ovaries. No differences in maturation rate, fertilization rate, cleavage rate and implantation rate were observed when oocytes from a polycystic ovary were compared with oocytes from an unstimulated normal ovary. In unstimulated cycles, the ovaries were grouped according to the presence of a dominant follicle. The incidence of apoptosis was significantly higher in granulosa cells from an ovary without a dominant follicle compared with granulosa cells from an ovary with a dominant follicle. The rates of maturation, fertilization and cleavage did not differ between the two groups.
本研究的目的是调查体外成熟(IVM)的未成熟人类卵泡颗粒细胞中凋亡的发生率,并比较(i)促卵泡激素(FSH)预处理和未预处理的正常卵巢之间以及(ii)多囊卵巢和正常卵巢之间凋亡颗粒细胞的发生率。此外,凋亡发生率与相应卵巢中卵母细胞的成熟、随后的受精和卵裂有关。纳入了70名接受70个IVM周期的女性。第1组由正常卵巢患者(n = 52)组成,第2组由多囊卵巢患者(n = 18)组成。第1组患者根据抽吸前是否用FSH预处理分为两组。在1a组(n = 27个周期),在未刺激的周期中获取卵母细胞。在1b组(n = 25个周期),在自发月经后第3天开始用重组FSH预处理3天后获取卵母细胞。在第2组中,所有患者在抽吸前均用重组FSH预处理3天。经阴道进行抽吸,将卵丘包裹的卵母细胞在受精前成熟28 - 30小时。从卵泡抽吸物中收集颗粒细胞。使用APOPTAG检测试剂盒对颗粒细胞进行染色并检测凋亡。与未预处理的正常卵巢卵泡以及FSH预处理的多囊卵巢卵泡相比,FSH预处理的正常卵巢卵泡中颗粒细胞的凋亡发生率降低。FSH预处理的多囊卵巢颗粒细胞与未刺激的正常卵巢颗粒细胞之间未发现差异。当将多囊卵巢的卵母细胞与未刺激的正常卵巢的卵母细胞进行比较时,未观察到成熟率、受精率、卵裂率和着床率的差异。在未刺激的周期中,根据优势卵泡的存在对卵巢进行分组。与有优势卵泡的卵巢颗粒细胞相比,无优势卵泡的卵巢颗粒细胞中凋亡发生率显著更高。两组之间的成熟率、受精率和卵裂率没有差异。