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Efficient use of oocytes by the perfusion of farm animal ovary

通过家畜卵巢灌注有效利用卵母细胞

基本信息

批准号：
62560267
负责人：
MIYAMOTO Hajime
金额：
$ 1.34万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560267/
关键词：
farm animal ovary ovarian function Graafian follicle ovulation perfusion medium ovarian histology ovarian histochemistry 培養装置グラーフ卵胞組織学ヤギ卵巣卵細胞の成熟 pH ブドウ糖

项目摘要

A technique for in vitro perfusion of goat ovaries is studied to use oocytes efficiently and to examine ovarian function. The effect of perfusion with different media on ovarian function and, in particular, on the phenomenon of follicular rupture and swelling is examined. A system has been developed for the perfusion of the goat ovary in vitro. The perfusion apparatus consisted of a recycling system with three major portions: an oxygenator, a roller pump, and a perfusion chamber. The functional integrity of the perfused ovary is confirmed by serial determinations of the perfusate pH, glucose and lactate concentrations, and by histology and histochemistry of the ovary.After several preliminary experiments, nine experiments, where the ovary containing Graafian follicles was used, were tried. In one of nine experiments the rupture of two Graafian follicles occurred by perfusion of the goat ovary. However in the other experiments the rupture of Graafian follicles did not occur. Perfusion o … More f the ovary caused swelling of Graafian follicles and small antral follicles, accompanied by a stromal edema. Graafian follicular rupture observed in perfusion system may be artifacts and careful selection of the perfusion medium is needed to avoid misinterpretation.Glucose consumption and lactate production occurred continuously during the 6-hour perfusion period. It was shown by light microscopy that almost normal ovarian structure was maintained throughout the perfusion, although interstitial edema formation was noted in the perfused ovaries. In the histochemical study, normal activity of 3beta-HSD and G6PDH was maintained in the theca interna and granulosa cells of follicles in the perfused ovaries. In contrast, electron microscopic observations indicated that the present perfusion system is not satisfactory to maintain ovarian tissue.In conclusion, a perfusion system of the farm animal ovary is useful for the efficient use of oocytes and for the study of ovarian function if perfusion technique could be improved. Less

研究山羊卵巢体外灌注技术，有效利用卵母细胞，检测卵巢功能。研究了不同介质灌注对卵巢功能的影响，特别是对卵泡破裂和肿胀现象的影响。建立了山羊卵巢体外灌注系统。灌注装置由氧合器、滚筒泵和灌注室三个主要部分组成的循环系统组成。灌注后卵巢的功能完整性可通过灌注液pH、葡萄糖和乳酸浓度的一系列测定以及卵巢的组织学和组织化学来证实。经过几次初步实验后，进行了9次实验，其中使用了含有Graafian卵泡的卵巢。在9个实验中的一个中，山羊卵巢灌注导致两个格拉芬卵泡破裂。然而，在其他实验中，没有发生毛囊破裂。卵巢灌注量增加，引起胃囊和小窦卵泡肿胀，伴间质水肿。在灌注系统中观察到的毛囊破裂可能是人为的，需要仔细选择灌注介质以避免误解。葡萄糖的消耗和乳酸的产生在6小时灌注期间连续发生。光镜下显示卵巢在整个灌注过程中基本保持正常结构，但灌注后卵巢出现间质性水肿。在组织化学研究中，灌注卵巢卵泡内膜和颗粒细胞中3β - hsd和G6PDH保持正常活性。电镜观察表明，目前的灌注系统不能很好地维持卵巢组织。综上所述，畜禽卵巢灌注系统的改进有助于卵母细胞的有效利用和卵巢功能的研究。少