Double vitrification of rat embryos at different developmental stages using an identical protocol

Theriogenology
Vladimir IsachenkoJose Luis Alabart

Abstract

The aim of the present investigation was to test the effectiveness of a method of vitrifying rat embryos at different stages of development (from early morula to expanding blastocyst) in a double vitrification procedure. Wistar rat embryos were vitrified and warmed in super-fine open-pulled straws (SOPS). Before being plunged into liquid nitrogen, the embryos were exposed to 40% ethylene glycol+0.75 M sucrose in TCM-199+20% fetal calf serum (FCS) for 20s at 38 degrees C. Subsequent warming and direct rehydration of the embryos was conducted in culture medium (TCM-199+20% FCS) at 38 degrees C. Early morula stage (7-10 blastomeres) embryos (n=358) were vitrified, warmed and cultured in vitro (EM group). Batches of these embryos were then cryopreserved again (revitrified) at the early blastocyst (EB group, n=87), blastocyst (B group, n=93) or expanding blastocyst stage (ExpB group, n=73). After the first (EM group) and repeated (EB, B, and ExpB groups) vitrification procedures, developmental rates of 81, 83, 34 and 76%, respectively were achieved (for EM-EB-ExpB P>0.1; for EM, EB, ExpB-B P<0.005). Our data demonstrate the possibility of using the described identical protocol for the SOPS vitrification of rat early morulae, early b...Continue Reading

References

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Citations

Jun 5, 2007·Theriogenology·C CuelloE A Martinez
Jul 16, 2010·Reproduction in Domestic Animals = Zuchthygiene·M A GilE A Martinez
Jul 15, 2015·Theriogenology·Emilio A MartinezMaria A Gil
Jul 13, 2005·Reproduction in Domestic Animals = Zuchthygiene·E A MartinezJ L Vazquez
Feb 9, 2018·Zoological Science·Sebastian J CharltonJulie Strand
Dec 7, 2021·Frontiers in Veterinary Science·Cristina CuelloEmilio A Martinez

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