In vitro analysis of the function in various premature follicles isolated enzymatically from rat ovaries.

对从大鼠卵巢酶法分离的各种早产卵泡的功能进行体外分析。

• 批准号: 62560294
• 负责人: ETOH Teiichi
• 金额: $ 1.28万
• 依托单位: University of Miyazaki
• 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 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560294/
• 关键词: follicle culture; Method of follicle separation; follicular development; follicnlar function; follicular steroid; PMSG; 新生児卵巣

A single follicle was isolated from rat ovary by digestion with enzymes and filtration with stainless steel wire gauze. The isolation was made use of the method applied in hamster by Roy c Greenwald. The isolated follicles were observed with phase-contrast or scanning-elect-ron microscopes and cultured individually for 4 days in two differentmethod in vitro, such as liquid medium or the same medium on agarose gel seat. The results are as follows. 1.The co-relation between diameter of follicles and the including ova was estimated. With increasing of the follicular diameter to 200 m, the ovular diameter increased to 70 m,but thereafter, the ovular diameter persisted in constant in spite of the follicular growth. The follicles were coated by basement membrane having many small projections, but shortly after the bigin-ing of incuvation, these were decoated the membrane and became a cluster of global cells. 2.Both preantral and small antral follicles in the liquid medium grew to monolayer c … More ellular seat holding cumulus cells and ovum in centre, and sometimes those cumulus cells changed the shape as similar as large antral follicle. And also, PMSG stimulated the changes and progest-erone production as well. 3. Follicles cultured in the medium on agarose gel seat did not grow clearly. This result indicated that the liquid was suitable for follicle incuvation than the agarose one. 4. The small follicles were relatively less sensitive to PMSG than the larger follicles. Ovaries from neonate rats, which were containing exclusively primary and preant-ral follicles, were placed in organ culture medium for 4 to 8 days in order to confirm vi-ability in in vitro circumstance. The follicles in these ovaries were maintained for as long time as 8 days histologically and produced a large amount of 4-androstene-3,17-dione, a precureser of estrogen, in limited period of 5 to 9 days in age. It is suggested that, from these experiments, enzymatically dissociated follicles even in neonate ovaries may be able to stimulate to growth in vitro and this incuvation system may be a useful tool for invest-igating very complex intraovarian endocrine mechanism. Less

通过酶消化并用不锈钢丝网过滤从大鼠卵巢中分离出单个卵泡。采用 Roy c Greenwald 在仓鼠身上应用的方法进行分离。用相差显微镜或扫描电子显微镜观察分离的卵泡，并在体外以两种不同的方法（例如液体培养基或琼脂糖凝胶座上的相同培养基）单独培养4天。结果如下。 1.估计卵泡直径与所含卵子的相关性。随着卵泡直径增加到200 m，卵子直径增加到70 m，但此后，尽管卵泡生长，卵子直径仍保持恒定。毛囊被基底膜覆盖，具有许多小突起，但在孵化后不久，这些基底膜就被脱去，并成为一簇整体细胞。 2.窦前卵泡和小窦卵泡在液体培养基中均生长成单层细胞座，中央含有卵丘细胞和卵子，有时这些卵丘细胞改变形状与大窦卵泡类似。而且，PMSG 还刺激了这些变化和孕激素的产生。 3.在琼脂糖凝胶座上的培养基中培养的卵泡没有明显生长。这一结果表明液体比琼脂糖更适合卵泡培育。 4.小卵泡对PMSG的敏感度相对较大卵泡较低。将仅含有初级卵泡和腔前卵泡的新生大鼠卵巢置于器官培养基中4至8天，以确认体外环境中的活力。这些卵巢中的卵泡在组织学上维持了长达8天的时间，并在5至9天的有限时间内产生大量的4-雄烯-3,17-二酮，一种雌激素的前体。这些实验表明，即使在新生儿卵巢中，酶解的卵泡也可能能够在体外刺激生长，并且该孵化系统可能是研究非常复杂的卵巢内内分泌机制的有用工具。较少的

项目成果

Teiichi, Etoh: "4-androstene-3,17-dione in the ovary , adrenal gland and peripheral blood plasma in neonate rats."

Teiichi, Etoh：“新生大鼠卵巢、肾上腺和外周血浆中的 4-雄烯-3,17-二酮。”

Noboru,Murakami.: Jap.J.Animal Reprod.32. 101-104 (1987)

村上登：Jap.J.Animal Reprod.32。

Noboru Murakami: Jap.J.Animal Reprod.32. 101-104 (1987)

村上登：Jap.J.Animal Reprod.32。

Noboru, Murakami: "A separation of intact preantral follicles from rat ovary by enzymatic dissociation and filtration" Jap. J. Animal Reproduction. 32. 101-104 (1987)

Noboru，Murakami：“通过酶解和过滤从大鼠卵巢中分离完整的腔前卵泡”Jap。

江藤禎一: 投稿準備中.

Teiichi Eto：准备提交。