Development of Effective Cryopreservation Method for Mouse Oocytes.
- Author:
Su Jin CHOI
;
Soo Kyung KIM
;
Ji Sun KIM
;
Jae Won CHO
;
Jin Hyun JUN
;
Hye Kyung BYUN
- Publication Type:Original Article
- Keywords:
Mouse oocyte;
Vitrification;
Slow-freezing;
Spindle;
Chromosome
- MeSH:
Animals;
Cryopreservation*;
Fluorescein;
Hand;
Metaphase;
Mice*;
Oocytes*;
Propidium;
Propylene Glycol;
Sucrose;
Survival Rate;
Vitrification
- From:Korean Journal of Fertility and Sterility
2004;31(1):75-81
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
OBJECTIVE: The purpose of this study was to evaluate the efficacy and effect of various cryopreservation method on the survival and the cytoskeletal stability of metaphase II mouse oocyte. METHODS: Mouse ovulated oocytes were collected and cryopreserved by a modified slow-freezing method with 1.5 M 1,2-propanediol (PrOH)+0.1 M sucrose or by vitrification using cryo loop and EM grid with 40% ethylene glycol+0.6 M sucrose. Four hours after thawing, intact oocytes were fixed and stained with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-beta-tubulin antibody to visualize spindle and propidium iodide (PI) to visualize chromosome. Spindle morphology was classified as follows: normal (barrel-shaped), slightly and absolute abnormal (multipolar or absent). RESULTS: Survival rate of the frozen-thawed oocytes in vitrification group was significantly higher than that of slow-freezing group (62.7% vs. 24.4%, p<0.01). Vitrification with cryo loop showed significantly higher survival rate than that with EM grid (67.7% vs. 53.5%, p<0.05). On the other hand, proportion of normal spindle and chromosome configurations of the frozen-thawed oocytes between two vitrification group was not significantly different. CONCLUSION: For mouse ovulated oocytes, vitrification with cryo loop may be a preferable procedure compared to slow-freezing method. Further study should be needed to investigate developmental competency of frozen-thawed mouse oocytes.
