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Effect of stage of development on survival of mouse embryos frozen-thawed rapidly

par Massip, Alban;Van der Zwalmen, P.;Leroy, Fernand
Référence Cryobiology, 21, 5, page (574-577)
Publication Publié, 1984

Article révisé par les pairs

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Résumé :

Embryos were recovered on Day 4 of pregnancy from superovulated random-bred OF1 Swiss albino mice. They were classified into four categories based on their stage of development: expanding blastocyst, blastocyst, early blastocyst, and compacted morula. They were then cooled at 2 °C/min from -7 to -25 °C in a freezing medium containing 1.36 M glycerol and 0.25 M sucrose in phosphate-buffered saline (PBS). At -25 °C, they were plunged into LN2 and thawed a few hours later in water at 20 °C. After washing in PBS, recovered embryos were cultured for 20 to 24 hr and the number of embryos that had developed normally was recorded. The results showed a clear effect of the stage of development on survival. Survival of expanding blastocysts and blastocysts was very low (1.4 and 21.8%, respectively) compared to that of early blastocysts and compacted morulae (69.4 and 73.5%). The more differentiated stage of the blastocyst (two kinds of cells) and the presence of a blastocoelic cavity may explain the differences observed under our cooling conditions. As a further test of viability, 93 blastocysts that had developed in culture for 20 hr from 153 frozen-thawed early blastocysts and compacted morulae (60.8%) were transferred to 8 recipient mice. Seven became pregnant, yielding 38 82 normal live young (46.3%). © 1984.