HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Keskintepe, L. Articles by Brackett, B. G. Search for Related Content PubMed PubMed Citation Articles by Keskintepe, L. Articles by Brackett, B. G. Agricola Articles by Keskintepe, L. Articles by Brackett, B. G.

Bovine Blastocyst Development from Oocytes Injected with Freeze-Dried Spermatozoa¹

^a Transgenic and Embryonic Stem Cell Core, IMMAG, Medical College of Georgia, Augusta, Georgia 30912 ^b Department of Gynecology, Medical College of Georgia, Augusta, Georgia 30912 ^c Department of Physiology and Pharmacology, College of Veterinary Medicine, The University of Georgia, Athens, Georgia 30602

Pronuclear formation, and the chromosomal constitution and developmental capacity of bovine zygotes formed by intracytoplasmic sperm injection with freeze-dried (lyophilized) spermatozoa were evaluated. Frozen-thawed spermatozoa were selected, freeze-dried, and stored at 4°C until use. After 22–24 h of in vitro maturation oocytes were denuded and injected singly with a lyophilized spermatozoon. Injected oocytes were activated by treatment with 10 µM ionomycin (5 min) alone and in combination with 1.9 mM 6-dimethylaminopurine (DMAP) for 4 h. Ionomycin plus DMAP activation treatment resulted in a significantly higher proportion of sperm-injected oocytes with two pronuclei than was found after activation with ionomycin alone (74% vs. 56%; P < 0.03). The rates of cleavage, morula, and blastocyst development of sperm-injected oocytes treated with ionomycin plus DMAP were higher than after activation with ionomycin alone (63.3%, 34.2%, and 29.6% vs. 44.7%, 18.7%, and 10.6%, respectively; P < 0.05). Seventy-three percent of blastocysts produced with lyophilized sperm were diploid. These results demonstrate that in vitro-matured bovine oocytes can be fertilized with freeze-dried sperm cells, and that resultant zygotes can develop into karyotypically normal blastocysts.

First decision: 15 June 2001.

¹ A portion of this work was presented at the 27th Annual Conference of the International Embryo Transfer Society, Omaha, NE, in 2001.

² Correspondence: Levent Keskintepe, Sher Institute for Reproductive Medicine, 3121 South Maryland Avenue, Suite 300, Las Vegas, NV 89109. FAX: 702 892 9666; lkeskintepe{at}sherinstitute.com

This article has been cited by other articles:

				K.-B. Lee and K. Niwa Fertilization and Development In Vitro of Bovine Oocytes Following Intracytoplasmic Injection of Heat-Dried Sperm Heads Biol Reprod, January 1, 2006; 74(1): 146 - 152. [Abstract] [Full Text] [PDF]

				I.-K. Kwon, K.-E. Park, and K. Niwa Activation, Pronuclear Formation, and Development In Vitro of Pig Oocytes Following Intracytoplasmic Injection of Freeze-Dried Spermatozoa Biol Reprod, November 1, 2004; 71(5): 1430 - 1436. [Abstract] [Full Text] [PDF]

				T. L. Nedambale, A. Dinnyes, X. Yang, and X. C. Tian Bovine Blastocyst Development In Vitro: Timing, Sex, and Viability Following Vitrification Biol Reprod, November 1, 2004; 71(5): 1671 - 1676. [Abstract] [Full Text] [PDF]