Vitrification of Mouse Blastocyst.
- Author:
Yu Jin LEE
1
;
Mi Young KIM
;
Seok Won LEE
;
Yu Il LEE
Author Information
1. Department of Obstetrics and Gynecology, Chonnam National University Medical School, Gwangju, Korea.
- Publication Type:Original Article
- Keywords:
Mouse blastocyst;
EFS40 solution;
VS solution;
Vitrification
- MeSH:
Animals;
Blastocyst*;
Dimethyl Sulfoxide;
Embryonic Structures;
Ethylene Glycol;
Ficoll;
Humans;
Mice*;
Sucrose;
Toxicity Tests;
Trehalose;
Vitrification*
- From:Korean Journal of Obstetrics and Gynecology
2004;47(7):1348-1354
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
OBJECTIVE: The aim of this study was to compare the survival and developmental rate of two vitrification solutions for the vitrification of mouse expanded blastocysts. METHODS: Mouse embryos were obtained at 2 cell stage and cultured to expanded blastocyst stage in Human Tubal Fluid (HTF) medium supplemented with 10% serum substitute supplement (SSS). The vitrification solutions used were EFS40 and VS. EFS40 consisted of 40% ethylene glycol, 18% ficoll, and 0.5 M sucrose while VS consisted of 20% ethylene glycol, 20% DMSO, and 10% 1,3-butanediol diluted in Dulbecco's phosphate-buffered saline (DPBS) medium supplemented with 10% calf serum (CS). Toxicity was tested by exposing expanded blastocysts to vitrification solution. The vitrification procedure used for EFS40 was performed in three steps, after which they were warmed in 5 steps with 0.5 M sucrose. VS was performed in two steps, after which they were warmed with 1.0 M trehalose. Recovery, survival and hatching rate per expanded blastocysts recovered were compared between two groups. RESULTS: In toxicity test, survival and hatching rate of EFS40 group were 95% and 100%, respectively. In contrast, survival and hatching rate of VS group were 100% and 87.5%, respectively. After vitrification and warming in solution, recovery rate for EFS40 group was 73.7% whereas recovery rate for VS group was 66.5%. After 24 h culture, survival and hatching rate were 80.5% and 20.7% for EFS40 and 66% and 0% for VS group, respectively. After 48 h culture, survival and hatching rate were 69% and 33.3% for EFS40 and 58.3% and 1.9% for VS group, respectively. Survival and hatching rate in EFS40 group were significantly higher than those found in VS group. CONCLUSION: The EFS40 solution was better than VS solution for vitrification of mouse expanded blastocysts.
