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Updated: Aug 8, 2026
Cryopreservation of Mouse Embryos by Ethylene Glycol-Based Vitrification
Published on: November 18, 2011
Low temperature preservation of mouse fetal germ cells at 4 degrees C
1Research Institute for Animal Developmental Biotechnology and Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, Japan, 631.
Abstract:
Mouse fetal germ cells(FGCs) isolated or within genital ridges from male fetuses at 15.5 d post coitum were preserved at 4 degrees C for 3-15 d with TCM-199 medium supplemented with 10, 50 or 100% fetal bovine serum (FCS) and 0.1, 0.6 or 1.0 M sucrose. The viability of FGCs was assessed by the dye exclusion test with trypan blue and nuclear transfer into enucleated oocytes and then into enucleated 2-cell embryos. Forty-one to 45 % of FGCs survived for 5 d after preservation in the medium containing 50% FCS and 0.1 M sucrose according to the results of the dye exclusion test. But significant good effect of sucrose was not observed regardless of its concentration. When genital ridges were preserved, 15-22% of FGCs survived even 10 d after preservation. The efficiency of the in vitro development of FGCs to blastocysts was similar when they were fused with enucleated oocytes and 2-cells blastomeres after 2 to 4 d storage(27-33%) compared with that of control FGCs(56%).
Insights
Preserving mouse fetal germ cells (FGCs) in a medium with 50% fetal bovine serum (FCS) and 0.1 M sucrose allowed 41-45% viability for 5 days. Sucrose concentration did not significantly impact survival, but preserving FGCs within genital ridges extended survival to 10 days.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cryobiology
Background:
- Mouse fetal germ cells (FGCs) are crucial for reproductive potential.
- Effective preservation methods for FGCs are needed for research and potential therapeutic applications.
Purpose of the Study:
- To investigate the viability and developmental potential of mouse FGCs after short-term preservation at 4°C.
- To evaluate the effects of fetal bovine serum (FCS) and sucrose concentrations on FGC preservation.
Main Methods:
- FGCs were isolated or maintained within genital ridges from male mouse fetuses (15.5 days post coitum).
- Preservation was conducted at 4°C for 3-15 days using TCM-199 medium with varying concentrations of FCS (10-100%) and sucrose (0.1-1.0 M).
- Viability was assessed using trypan blue dye exclusion and nuclear transfer into enucleated oocytes and 2-cell embryos.
Main Results:
- 41-45% FGC viability was observed after 5 days of preservation in medium with 50% FCS and 0.1 M sucrose.
- Sucrose concentration did not show a significant beneficial effect on FGC viability.
- FGCs preserved within genital ridges demonstrated 15-22% survival after 10 days of preservation.
- In vitro development to blastocysts after 2-4 days of storage showed similar efficiency (27-33%) when fused with oocytes/blastomeres compared to controls (56%).
Conclusions:
- Short-term cold preservation of mouse FGCs is feasible, with optimal conditions identified for initial viability.
- Preservation within the genital ridge structure enhances FGC survival duration.
- While some viability is maintained, developmental potential to blastocysts is reduced after preservation and nuclear transfer.
