着床近時の家兎胞胚のin vitro培養について〔英文〕

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Rabbit blastocysts explanted at 6 day and 18 hr post coitum were cultured in vitro, and the effects of volume and surface area of the medium exposed to gas phase on the embryonic development were examined.Three types of culture vessel; Carrel's flask (vol.; 180 & 90cm3, medium vol.; 48 & 24ml, medium surface; 12 & 6cm2), Earl's flask (vol.; 12cm3, medium vol.; 9.6ml, medium surface; 2.4cm2) and test tube (vol.; 11.8 cm3, medium vol.; 6.3ml, medium surface; 1.77cm2) containing medium 199 supplemented with 20% calf serum and 5% Difco'bacto-tryptose phosphate' were employed. The proportion of the embryos developed to the stage more than 10 somites varied with the type of culture vessels. The proportion tended to increase with the increase of medium volume and of area of medium surface facing to gas phase. In 72 hr incubation with the Carrel's flask (180cm3), 32 (70%) of 46 embryos tested reached the stage of somites with neural fold, and 19 (40%) of them proceeded to more than 10 somites. Further cultivation beyond 3 days led to malformation in many cases. The most advanced development attained was the stage of 20 somites with cephalic subdivision, optic vesicles and pulsating heart, which compares with normal 9.5 day embryo. In the smaller culture vessels (Earl's flask & Test tube), 25% of the embryos tested developed to the stage of somites but many other specimens collapsed after reaching to the primitive streak stage. The poorest growth was found in the embryos cultured in Earl's flask and Test tube kept in inclined position (30°). Many of the embryos in these inclined vessels collapsed without attaining at primitive streak stage. The failure in development seemed to be due to a friction between the surface of blastocyst and the wall of vessel.These results indicate that some of basic factors which affect in vitro embryonic growth of rabbit from blastocyst to somite stage are; (1) medium-volume, (2) area of medium surface facing to gas phase and (3) mechanical stimuli to embryonic area. A large amount of medium and a wide surface area of medium exposed to gas phase are neccessary conditions for cultivation of rabbit embryo to allow the development from blastocyst to somite stage. The mechanical stimuli to embryonic disc should be avoided for cultivation of embryo at blastocyst stage, since a trivial mechanical contact to the wall of culture vessel may disturb the continued embryonic development.
社団法人日本畜産学会の論文

着床近時の家兎胞胚のin vitro培養について〔英文〕

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