Capacitation-like membrane changes and prolonged viability in vitro of equine spermatozoa cultured with uterine tube epithelial cells

J. E. Ellington From the Department of Clinical Sciences, New York State College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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B. A. Ball From the Department of Clinical Sciences, New York State College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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B.J. Blue From the Department of Clinical Sciences, New York State College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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C. E. Wilker From the Department of Clinical Sciences, New York State College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Summary

Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (utec)-conditioned medium, and direct culturing of spermatozoa with utec (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with utec-conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P. < 0.05) capacitation-like changes than did the control, a modified Tyrode’s medium. More (P < 0.05) spermatozoa were viable after 24 hours of utec coculturing than in the control incubation.

Summary

Reliable capacitation of equine spermatozoa has been a major obstacle in the development of equine in vitro fertilization. Experiments were done to compare in vitro capacitation of equine spermatozoa by use of heparin/caffeine, calcium ionophore, uterine tube epithelial cell (utec)-conditioned medium, and direct culturing of spermatozoa with utec (coculturing). Capacitation-like changes, as determined by chlortetracycline membrane staining patterns, developed with utec-conditioned medium and coculturing, equivalent to that with calcium ionophore. Both of these treatments induced more (P. < 0.05) capacitation-like changes than did the control, a modified Tyrode’s medium. More (P < 0.05) spermatozoa were viable after 24 hours of utec coculturing than in the control incubation.

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