Optimized Method For Bovine Blastocyst Vitrification Using A Simple Hand-Made Cryotip
نویسندگان
1 Faculty of Basic Science, Islamic Azad University, Marvdasht Branch, Marvdasht, Iran
2 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
3 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
4 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
5 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
6 Anatomy Department, Isfahan University of Medical Sciences, Isfahan, Iran
7 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
8 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
9 Anatomy Department, Isfahan University of Medical Sciences, Isfahan, Iran
10 Anatomy Department, Isfahan University of Medical Sciences, Isfahan, Iran
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چکیده
Objective: This study introduced a simple method for bovine blastocyst vitrification.Materials and Methods: Bovine blastocysts were produced in vitro by means of a whole co-culture system with vero cells. The blastocysts were randomly divided 1:3 into either vitrification (100 blastocysts) or control (43 blastocysts) groups. For vitrification,expanded - blastocysts were incubated first in equilibration medium for 8 minutes and then in the vitrification solution for 1 minute. The blastocysts were then loaded in the tip of a handmade cryotip for immediate - deep freezing in liquid nitrogen. Frozen embryos were then warmed by directly immersing the tips in sequential warming solutions . warmed embryos were cultured for a further period of 48 hours when the ratios of re-expansion, hatching and degeneration were compared with the control group.Results: After warming, in the vitrified and control groups the ratios of re-expansion were 78.5% ± 0.067 and 81.6% ± 0.072, the ratios of hatching were 43.7% ± 0.083 and 49.8% ± 0.089 and the ratios of degeneration were 36% ± 0.082 and 22.3% ± 0.087, respectively, which were not significantly different between the two groups.Conclusion: Post - warming survival of the vitrified and non - vitrified embryos were not significantly different, handmade cryotips can be used as an efficient and feasible device for bovine blastocysts vitrification.
