Factors affecting the survival of frozen-thawed mouse spermatozoa.
Mouse epididymal spermatozoa were frozen in solutions containing various compounds with different molecular weights, and the factors affecting the postthawing survival were examined. Monosaccharides (glucose, galactose) had almost no protective effect regardless of the concentration and the temperature of exposure. On the other hand, disaccharides (sucrose, trehalose) and trisaccharides (raffinose, melezitose) resulted in higher survival rates, especially at a concentration of around 0.35 mol/kg H(2)O (0.381-0.412 Osm/kg). Macromolecules, such as PVP10, Ficoll 70, bovine serum albumin, and skim milk had almost no effect, but compounds with a molecular weight of about 800, such as metrizamide and Nycodenz, had some protective effect. When a raffinose solution was supplemented with 10% metrizamide, resulting in an osmolality of approximately 0.400 Osm/kg, a high survival rate was obtained. Solutions at about 0.400 Osm/kg containing trehalose alone, trehalose + metrizamide, raffinose alone, and raffinose + metrizamide, were all effective for sperm freezing; frozen-thawed sperm could fertilize oocytes, and the resultant embryos could develop to live young after transfer. For freezing mouse spermatozoa, aqueous solutions at approximately 0.400 Osm/kg containing a disaccharide or a trisaccharide seem to be effective.
An
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TZ
Laboratory of Animal Science, College of Agriculture, Kochi University, Nankoku, Kochi, 783-8502, Japan.
Iwakiri
M
M
Edashige
K
K
Sakurai
T
T
Kasai
M
M
D000818:Animals; D002241:Carbohydrates; D002417:Cattle; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D005260:Female; D005307:Fertilization in Vitro; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009994:Osmolar Concentration; D011887:Raffinose; D012662:Semen Preservation / Q000379:methods*; D013094:Spermatozoa / Q000166:cytology; D014199:Trehalose
D002241:Carbohydrates; D003451:Cryoprotective Agents; D014199:Trehalose; D011887:Raffinose