A mechanical and enzymatic approach to recovery of ovarian follicular oocytes in rabbits.
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概要
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In order to investigate the morphology of oocytes which were recovered from ovarian follicles at several developmental stages and to examine the <I>in vitro</I> maturity of recovered oocytes, rabbit ovarian oocytes were recovered by mincing the ovary with a pair of razors and/or cutting isolated follicles. Twenty four point six oocytes per ovary were recovered by mincing the ovary with a pair of razors, and 19.9% of them were surrounded by a thick layer of cumulus cells. The mass of ovarian tissue remnant was digested by a collagenase solution to isolate ovarian follicles at several developmental stages. A great number of ovarian follicles (304.9±114.0 per ovary) could be collected by enzymatic isolation, however, it is difficult to isolate follicles over 700μm in diameter. The oocytes from antral follicles which were isolated enzymatically ( ?? 300 Am in diame-ter) were obtained by cutting with a pair of razors, but oocytes could not be recovered from preantral follicles by the same procedure. Some oocytes obtained by cutting follicles were cultured for 20 hr in TCM 199 supplemented with 15% FBS for <I>in vitro</I> matu-ration. Oocytes with a thick layer of cumulus cells were obtained only from follicles over 700 Am in diameter. They emitted the first polar body <I>in vitro</I> at a higher rate as compared with the other types of oocytes, which were surrounded by a thin or partial cumulus cell layer or had no cumulus cells at all. All types of oocytes resumed meiosis <I>in vitro</I>, although high percentages of the oocytes from the smaller follicles and/or in the cumulus free oocytes degenerated. These results suggest that a combination of mincing ovaries and cutting follicles which were isolated enzymatically or mechanically is useful for recovering a great number of oocytes. However, <I>in vitro</I> maturity of those oocytes depends on the follicular size and/or cumulus cells' adhesion to the oocytes.
- 日本繁殖生物学会の論文
日本繁殖生物学会 | 論文
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