Perfusion of the rat ovary in vitro: methodology, induction of ovulation, and pattern of steroidogenesis.
Perfusion of the rat ovary in vitro: methodology, induction of ovulation, and pattern of steroidogenesis.
复制标题
大鼠卵巢体外灌注：方法、排卵诱导和类固醇生成模式。
DOI：
10.1095/biolreprod30.5.1135
复制
发表时间：
1984
影响因子：
3.6
通讯作者：
W. L. Le Maire
中科院分区：
文献类型：
--
作者：
R. Koos;F. J. Jaccarino;R. Magaril;W. L. Le Maire
建立了一种体外灌注大鼠卵巢的方法，并对其进行了表征。未成熟大鼠（27-29日龄）用20 IU孕马血清促性腺激素（PMSG）致敏。两天后进行剖腹手术，主动脉和腔静脉插管，分离右侧卵巢，结扎除右侧卵巢动脉和静脉外与主动脉和腔静脉相连的所有血管。将制备物置于灌注装置中，并用含4%牛血清白蛋白的含氧培养基199（37 ℃，pH 7.4）灌注长达20 h。灌注压70-90 mm Hg，平均流量1 ml。min .卵巢通过从装置中回收卵母细胞确认排卵。9个卵巢在没有进一步处理的情况下进行灌注（对照）;该组中仅发生1次排卵。9个卵巢接受促黄体生成激素（LH; 0.1微克/毫升）1小时后开始灌注，71排卵（范围，3-13/卵巢）。LH给药后9 h前未开始排卵。频繁采集培养基样品用于测量孕酮、雌二醇和雄烯二酮。所有三种类固醇的水平迅速上升，显着响应LH，但仅略有增加，在控制灌注。本研究证实了大鼠卵巢在体外可诱导排卵，为进一步研究排卵机制提供了依据。
A method for perfusion of the rat ovary in vitro for the study of ovulation was developed and characterized. Immature rats (27-29 days old) were primed with 20 IU of pregnant mare's serum gonadotropin (PMSG). Two days later a laparotomy was performed, the aorta and vena cava were cannulated, the right ovary was isolated, and all vessels connecting with the aorta and vena cava except for the right ovarian artery and vein were ligated. The preparation was placed in the perfusion apparatus and perfused for up to 20 h with oxygenated Medium 199 containing 4% bovine serum albumin (37 degrees C, pH 7.4). The perfusion pressure was 70-90 mm Hg and the average flow was 1 ml . min . ovary. Ovulation was confirmed by oocyte recovery from the apparatus. Nine ovaries were perfused without further treatment (controls); only 1 ovulation occurred in this group. Nine ovaries received luteinizing hormone (LH; 0.1 microgram/ml) 1 h after the start of perfusion; 71 ovulations resulted (range, 3-13/ovary). Ovulations did not begin before 9 h after LH administration. Samples of medium were taken frequently for measurement of progesterone, estradiol and androstenedione. Levels of all three steroids rose rapidly and markedly in response to LH but increased only slightly in control perfusions. This study demonstrates that ovulation in the rat ovary can be induced in vitro and provides a basis for further studies on the mechanism of ovulation.