Studies on deep-freezing of boar semen. IV. Additional effects of DMSO as a protective agent
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As a kind of protective agent against freezing-damage of animal cells, dimethyl sulfoxide (DMSO) was compared with glycerol concerning boar sperm. The semen was collected by means of artificial vagina from the 3 healthy boars. The basic diluent was composed of the 20% egg yolk SGS solution (IIDA& ADACHI: 1966).<BR>At the first trial, seminal plasma was replaced by the 1st solution, with no protective agent included, using a centrifugator at a room temperature of 25°C. The material was cooled from the room temperature to 10°C at the rate of 1°/16 min. At 10°C, this material was added with the same volume of 2nd solution which contained various percentages of protective agent. After equilibration at 10°C for 2 hours, the material was frozen to -79°C at the rates of 1°/min from 10°C to -20°C and of 5°C/min from -20°C to the final temperature.<BR>The results are summarized as follows:<BR>1) The evaluation of toxic effect of DMSO (5, 7, 10 and 20<I>%</I> v/v) at the stage of pre-freezing was found to be equal to the toxic effect of 7% glycerol at 10°C within 6 hours.<BR>2) In the observation of freeze-thawed sperm motilities, 1, 5, 10 and 20 % DMSO showed some protection of sperm cells against freezing damage, however they were less effective than that of 7% glycerol level.<BR>3) Within those concentrations of DMSO used for this experiment, 5% level was the most effective for sperm motility.<BR>At the second trial, the 2nd solution (2, 5, 10 and 15% DMSO and 14% glycerol) was added as described above but at 5°C, and then the material was frozen in pellet form (NAGASE & NIWA : 1964) at once.<BR>4) From the above experimental result it was revealed that the order of effective concentration of DMSO was 5, 2.5, 1 and 7.5% in the final levels, however, there was no levels that were superior to 7% glycerol.<BR>5) There are no significant diflerences analized respectively between the control and 5% and 2.5% DMSO levels, and 7.5% level showed highly significant difference between the other levels (Fig. 2).<BR>From these results, the available percentage of DMSO on freezing of boar sperm was approximately 5%.
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