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Survival of mouse morulae and blastocysts derived from in vitro fertilization after ultra rapid freezing
1Nippon Institute for Biological Science, Tokyo, Japan.
Abstract:
Mouse morulae and blastocysts derived from in vitro fertilization were placed in a highly concentrated vitrification solution (DAP 213:2 M dimethyl sulfoxide, 1 M acetamide, 3 M propylene glycol in PB 1) in a sampling tube at room temperature, plunged into liquid nitrogen within ten seconds and cryopreserved. Thawing was carried out by directly pouring 37 degrees C 0.3 M sucrose solution into the sampling tube. The ratios of morphologically normal embryos at the morula and blastocyst stages after thawing were 92.0% (149/162) and 13.3% (13/98), respectively. The rate of development from morphologically normal morulae into blastocysts in vitro was 83.1% (74/89). Some of the morphologically normal morulae were transferred to pseudopregnant recipients immediately after thawing, and 45.0% (27/60) of the embryos developed into normal young.
Insights
Cryopreservation of mouse embryos using a novel vitrification solution (DAP 213) achieved high survival rates for morulae. This method enables successful development into viable offspring after thawing, offering a promising technique for embryo preservation.
Area of Science:
- Reproductive Biology
- Cryobiology
Background:
- In vitro fertilization (IVF) generates embryos requiring preservation.
- Vitrification is a key cryopreservation technique for embryos.
Purpose of the Study:
- To evaluate the efficacy of a new vitrification solution (DAP 213) for mouse embryo cryopreservation.
- To assess the post-thaw viability and developmental potential of mouse morulae and blastocysts.
Main Methods:
- Mouse morulae and blastocysts were vitrified using DAP 213 solution (2M DMSO, 1M acetamide, 3M PG) at room temperature.
- Rapid plunging into liquid nitrogen for cryopreservation.
- Thawing involved direct addition of 37°C 0.3M sucrose solution.
Main Results:
- High survival rate of 92.0% for morphologically normal morulae post-thaw.
- Lower survival rate of 13.3% for morphologically normal blastocysts post-thaw.
- 83.1% of surviving morulae developed into blastocysts in vitro, and 45.0% resulted in live offspring after transfer.
Conclusions:
- The DAP 213 vitrification solution is effective for cryopreserving mouse morulae with high survival rates.
- This method supports successful embryonic development and live birth, indicating its potential for assisted reproductive technologies.