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https://hdl.handle.net/2142/19579
Description
Title
Storage of mouse embryos of different genotypes
Author(s)
Rosario, Carlos Julio
Issue Date
1994
Doctoral Committee Chair(s)
Graves, Charles N.
Department of Study
Animal Sciences
Discipline
Animal Sciences
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Genetics
Biology, Cell
Agriculture, Animal Culture and Nutrition
Language
eng
Abstract
The purpose of this study was to determine if either the embryo or maternal genotype contribute to the difference in the percentage survival of embryos after cooling and storage. A total of 2669 eight-cell mouse embryos of different genotypes (ICR, C57BL/6n, ICR x C57 and C57 x ICR) were stored for one day at 4$\sp{\underline\circ}$C or at $-$196$\sp{\underline\circ}$C after vitrification or after slow cooling. Following storage the embryos were washed and cultured at 37$\sp{\underline\circ}$C under an atmosphere of 5% CO$\sb2$ in air and the percentage of embryos which developed to the blastocyst stage was assessed. The embryos stored at 4$\sp{\underline\circ}$C were stored in either phosphate buffeted saline (PBS) or PBS supplemented with sucrose, glycerol or ethylene glycol. The embryos stored in medium containing sucrose resulted in the highest percentage survival whereas embryos stored in medium containing ethylene glycol had the lowest percentage survival. For embryos stored in PBS, the highest embryo survival ratio was for the ICR embryos and the lowest was for the C57 embryos while the embryo survival for the ICR x C57 embryos was higher than for the C57 x ICR embryos. When sucrose was present in the storage medium, no differences were observed in the percentage of embryos developing to the blastocyst stage among the embryos of different genotypes. In medium containing either glycerol or ethylene glycol, the embryo survival percentage was not different for the ICR and C57 embryos, however the ICR x C57 embryos had a higher survival percentage than the C57 x ICR embryos. When embryos were vitrified in a solution containing 25% glycerol and 25% 1,2 propanediol in PB1, the ICR embryos had the highest survival percentage (57.2%) which was significantly different from the other three embryo genotypes, while the C57 embryos resulted with the lowest embryo survival (42.9%). The ICR x C57 and the C57 x ICR embryos resulted with 52.3 and 49.8% survival rates respectively which were not different (P $\le$ 0.05). When the embryos were slow cooled in 10% glycerol in PB1, stored at $-$196$\sp{\underline\circ}$ PC and thawed by the three step procedure, significant differences (P $\le$ 0.05) were observed in survival between the ICR (70.8%) and C57 (60.2%) genotypes. The ICR x C57 had 69.9% survival, which was significantly higher (P $\le$ 0.05) than the C57 x ICR (50.7%) embryos. Transfer of blastocysts stage embryos of the four genotypes to the uteri of day 4 pseudopregnant ICR and C57 recipients resulted in a lack of significant difference in either the pregnancy rate or the number of fetuses among recipients. (Abstract shortened by UMI.)
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