J. Anim Sci.
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J. Anim Sci. 1984. 59:454-459.
© 1984 American Society of Animal Science
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Characteristics of Bovine Spermatozoa after Migration through a Bovine Serum Albumin Gradient1

Lydia M. White2,3,, W. E. Beal3,4,, J. H. Bame5, R. G. Saacke5 and C. E. Marshall6

Virginia Polytechnic Institute and State University, Blacksburg 24061 and Select Sires, Inc., Plain City, OH 43064

Abstract

Experiments were conducted to determine the efficiency with which viable, morphologically normal bovine spermatozoa could be isolated using a discontinuous bovine serum albumin (BSA) gradient. In the first experiment, extended semen was layered on top of a BSA gradient (4% BSA over 10% BSA) contained in a 500-ml separatory funnel. When comparing 1,3, 5, 7, 14 or 21 x 109 spermatozoa applied to the gradient, the percentage of spermatozoa recovered from the lower third of the 10% BSA ranged from 2.9 to 18.5%. The greatest recovery was achieved when 1 x 109 sperm cells were applied. Increasing the number of spermatozoa applied to the gradient increased the percentage of spermatozoa remaining in the upper portions of the gradient. Motility of spermatozoa immediately after collection from the 10% BSA layer of the gradient was >90%, regardless of the number of spermatozoa applied. In a second experiment with freeze-thawed separated or unseparated spermatozoa, post-thaw motility (>60%) and acrosomal integrity (>85%) of separated spermatozoa (4 or 10% BSA layer) was greater (P<.05) than that of unseparated spermatozoa (38 and 66%, respectively). The discontinuous gradient excluded decapitated spermatozoa and spermatozoa with mid-piece and principal piece abnormalities from entering the lower layers. Sperm cells with head abnormalities were not separated. These data indicate that a population of spermatozoa with a high frequency of viable, motile, morphologically-normal bovine spermatozoa can be isolated using a discontinuous BSA gradient.

Footnotes

1 Funded in part by a grant from the Union Fork and Hoe Co. Foundation, Columbus, OH.

2 Research completed as partial requirement for a Master of Science degree at Virginia Polytechnic Institute and State Univ.; present address Division of Animal Science, West Virginia Univ., Morgantown 26S06.

3 Dept. of Anim. Sci., Virginia Polytechnic Institute and State Univ.

4 To whom reprint requests should be sent.

5 Dept. of Dairy Sci., Virginia Polytechnic Institute and State Univ.

6 Select Sires, Inc.






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Copyright © 1984 by the American Society of Animal Science.