In-vitro-Maturation porciner Oozyten auf Feederlayer ... - Dragon IVF

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In-vitro-Maturation porciner Oozyten auf Feederlayer ... - Dragon IVF

division was determined (n = 109 - 139 oocytes/group). Using the same test situation as

in Experiment 3, more oocytes were matured and fertilised. Those test groups without

the anti-inhibin antibody which had shown a poor result in Experiment 3 showed in this

new test series a tendency for a higher rate of division (32%, 32% and 34%). Those

oocytes grown on media with the lowest concentrations of anti-inhibin exhibited an

equivalent rate of division to this. In comparison, the group which had exhibited the

highest maturation rate in Experiment 3 - the group grown on the medium containing

only 10% pFF - now showed the lowest rate of division (25%). As the test situation for

these two experiments were the same, this result indicates that there is a great

discrepancy between the degree of nuclear and cytoplasmic maturation induced by the

various additives to the basic culture medium.

The degree of cytoplasmic maturation of the oocytes was then assessed in Experiment 6

(n = 158-220 oocytes/group). The test situations in this experiment, whereby the oocytes

were cultured with a feeder layer mirrored those set up in experiment 4. The best rates

of division occurred in the culture with 9% pFF, 1% anti-inhibin and a BRL feeder layer

(38%). The worst results occurred in the culture without either a protein source or a

feeder layer. The rate of division of all the other test groups lay between 25 and 32%.

In Experiment 7, a complex system of judgement for the classification of the test groups

was used (n = 84-137 oocytes/group). The rate of division, the integrity of the cytoplasm

and the number of nuclei after in vitro fertilisation were assessed for every oocyte. The

design of this experiment was the same as for Experiment 6. The worst results for all

three criteria were found in those test groups without a feeder layer. A cultivation

constellation of 9% pFF, 1% anti-inhibin and a BRL feeder layer produced the best

results for the rate of division, vitality and the percentage of zygotes with 2 or more

nuclei (39%, 89% and 40%, resp.). The cultures with 9% pFF, 1% anti-inhibin and pEIL

had a 35% rate of division, 94% vital oocytes and 50% fertilised oocytes containing 2 or

more nuclei. These two methods of cultivation can both, therefore, be recommended for

the in vitro maturation of porcine oocytes.

Experiment 8 considered the use of a feeder layer not only for the maturation phase but

also for the subsequent culture of the embryo (n = 68-80 oocytes/group). The best results

in both culture phases were obtained with a BRL feeder layer system (66% division rate,

84% vital oocytes and 78% oocytes with 2 or more nuclei). In comparison, the maturation

and embryo cultures without feeder layers exhibited a rate of division of 18%, 48% vital

oocytes, and 25% oocytes with 2 or more nuclei. As in comparison to the feeder-layersupported

embryo cultures, there were significantly poorer results for all three

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