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The use of sodium hyaluronate in freezing media for bovine and murine embryos

, : The use of sodium hyaluronate in freezing media for bovine and murine embryos. Cryobiology 30(2): 172-178

Four experiments were designed to determine whether sodium hyaluronate (SH) may be used to replace newborn calf serum (NSC) in murine and bovine embryo freezing media. A total of 780 mouse and 178 cattle embryos were frozen. Freezing media were prepared in Dulbecco's phosphate-buffered saline supplemented with 20% NCS and 10% glycerol (v/v) (medium A). In experiments I and II, NCS was replaced by 0.1% SH (w/v) in medium B and by 0.1% polyvinyl alcohol (PVA) (w/v) in medium C. In experiments III and IV, NCS was replaced in mouse embryo freezing medium by three different molecular weights of SH as follows: 0.1 or 0.2% (SH-1; lt 3 times 10-5 Da) in medium B, by 0.1 or 0.05% (SH-2; 5-7.5 times 10-5 Da) in medium C, and by 0.1 or 0.025% (SH-3; gt 1.2 times 10-6 Da) in medium D. Embryos were frozen and thawed using standard procedures. Glycerol was diluted from embryos after thawing in a single step with 1.0 M sucrose. After 48 h in culture (Ham's F-10 medium for cattle embryos and BWW medium for mouse embryos), embryos were evaluated for development to expanded or hatched blastocysts. Survival rates were compared by chi-2 analysis. In experiment I, there were no significant differences in mouse embryo survival or development between NCS, SH, and PVA groups (84.1, 79.2, and 83.3%, respectively). In experiment II, there were no significant differences in bovine embryo survival between NCS and SH groups (67.2 and 67.7%), but both had significantly higher survival than the PVA group (37.9%; P lt 0.05). In experiment III, there was a significantly lower (P lt 0.05) embryo survival rate in the low-molecular-weight SH-1 group (18.6%) than in medium (SH-2)- or high (SH-3)-molecular-weight groups (78.6 and 92.5%, respectively). Although doubling the concentration of SH-1 in experiment IV resulted in improved embryo survival (63.4%), survival rates were significantly higher (P lt 0.05) in the medium-molecular-weight (SH-2; 82.3%) and high-molecular-weight (SH-3; 86.0%) groups despite reduced concentrations of these compounds in the freezing medium. Results suggest that SH can be used to replace biological sera in embryo freezing media.

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Accession: 002530604

PMID: 8319487

DOI: 10.1006/cryo.1993.1016

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