Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print June 23, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.031260
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BIOLOGY OF REPRODUCTION 71, 1430–1436 (2004)
DOI: 10.1095/biolreprod.104.031260
© 2004 by the Society for the Study of Reproduction, Inc.

Reproductive Technology

Activation, Pronuclear Formation, and Development In Vitro of Pig Oocytes Following Intracytoplasmic Injection of Freeze-Dried Spermatozoa1

In-Kiu Kwon3, Ki-Eun Park3, and Koji Niwa2,4,3

The Graduate School of Natural Science and Technology,3 Okayama University, Okayama 700-8530, Japan Department of Animal Science and Technology,4 Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan

The fertilization of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa was evaluated. Activation and male pronuclear (MPN) formation were better in oocytes injected with isolated freeze-dried sperm heads than whole freeze-dried spermatozoa, but cleaved embryos were generally difficult to develop to the morula or blastocyst stage. When spermatozoa were freeze-dried for 24 h, oocyte activation and MPN formation in activated oocytes after sperm head injection were inhibited. Embryo development to the blastocyst stage was only obtained after injecting sperm heads isolated from spermatozoa freeze-dried for 4 h and stored at 4°C. The proportion of embryos that developed to the blastocyst stage was not increased by the treatment of injected oocytes with Ca ionophore (5–10 µM). Increasing the sperm storage time did not affect oocyte activation or MPN formation, but blastocyst development was observed only after 1 mo of storage. These results demonstrate that pig oocytes can be fertilized with appropriately freeze-dried spermatozoa and that the fertilized oocytes can develop to the blastocyst stage.

1 Supported in part by a Grant-in-Aid for Creative Scientific Research from the Japan Society for the Promotion of Science (13GS0008).

2 Correspondence. FAX: 81 86 251 8388; kniwa{at}cc.okayama-u.ac.jp






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Copyright © 2004 by the Society for the Study of Reproduction.