Potential of two-cell mouse embryos to develop to term despite partial damage after cryopreservation.
Approximately 18% of cryopreserved 2-cell mouse embryos of 26 different batches showed various degrees of morphological damage after the freeze-thaw process. Normal and damaged morphology were assessed by light microscopy and the ability of an embryo to develop in vitro to a blastocyst, or to develop to term, after transfer to foster mothers. Using vital stains such as Fluorescein-diacetate (FDA) and 4', 6-Diamidino-2-Phenylindole (DAPI) it was found that in approximately 82% of the cases, both of the 2 blastomeres of the cryopreserved embryos survived the freeze-thaw process; in 10% only one cell survived the process; and in 8% none survived. Normally, only intact 2-cell embryos are considered for transfer. Here it was shown that over 60% of the partially damaged embryos developed in vitro to the blastocyst stage and, of those, 26% developed to term after transfer to suitable foster mothers. Although the inner cell mass (ICM) appeared to remain smaller during culture after the transfer of partially damaged 2-cell stage embryos, no difference during gestation period was found compared with intact embryos.
Rülicke
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Central Biological Laboratory, University Hospital of Zürich, Switzerland.
Autenried
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D000818:Animals; D001757:Blastomeres / Q000166:cytology*; D004396:Coloring Agents; D015925:Cryopreservation / Q000662:veterinary*; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005260:Female; D005452:Fluoresceins; D066298:In Vitro Techniques; D007211:Indoles; D051379:Mice; D011247:Pregnancy; D051381:Rats
D004396:Coloring Agents; D005452:Fluoresceins; D007211:Indoles; C007293:DAPI; C018506:diacetylfluorescein