Long-term storage of mouse embryos at —196 °C: the effect of background radiation

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Abstract
SUMMARY: In order to test the feasibility of preservation, of genetic stocks of mice by storage in liquid nitrogen, mouse embryos at the 8-cell stage, were frozen and stored in liquid nitrogen at – 196 °C under increased radiation exposures of 1·8×, 9× and 84× background levels for periods of 6–8 months, 10–12 and 27–29 months, the 1·8 × level being regarded as a control. Their survival rates to the blastocyst stage, and after transfer to recipient females, to foetal or liveborn stages were then compared with those of unfrozen or short term frozen control embryos. The freezing processper secaused a marked loss of viability, in comparison with the unfrozen controls, but at the 1·8× radiation level there was then no further loss in viability even at the longest storage time (27–29 months). Similarly, at the 9× radiation level there was no loss of viability during storage up to 29 months, but at the 84× level the proportions of implanted embryos and live foetuses were slightly reduced. It was not clear if this was a true effect of radiation, since it was not related to time of storage. Considering all groups, about 20–30% of the embryos originally frozen were recovered as foetuses or liveborn young. It is concluded that the preservation of genetic stocks by storage in liquid nitrogen is a feasible proposition.