Development of pig fetuses from embryos frozen at -35.DEG.C.

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The survival of pig embryos after freezing at -35°C has been investigated. Expanding to hatched blastocysts were recovered from a Hampshire sow 6 days after the artificial insemination with semen from a Duroc boar. The recovered embryos were frozen in a Dulbecco's PBS containing 1.5M glycerol and 20% fetal calf serum. The embryos were cooled at 1°C/min from 35°C to -6.8°C, kept at -6.8°C for 10 min while seeding was accomplished, and then they were cooled at 0.3°C/min from -6.8°C to -35°C. When the embryos reached -35°C, they were immediately thawed by direct transfer to a 37°C water bath for about 20 seconds. Thirteen of 14 embryos (93%) showed normal development after incubation for 18 hrs. Two embryos stained with fluorescein diacetate (FDA) revealed sufficient accumulation of intracellular fluorescein. The recipient gilt which had received 11 embryos became pregnant with plural fetuses revealed by ultrasonic echography at 70 days after embryo transfer.
日本繁殖生物学会の論文