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1 August 2002 Bovine Blastocyst Development from Oocytes Injected with Freeze-Dried Spermatozoa
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Abstract

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.

Levent Keskintepe, Gabriela Pacholczyk, Anna Machnicka, Karen Norris, M. Akif Curuk, Iqbal Khan, and Benjamin G. Brackett "Bovine Blastocyst Development from Oocytes Injected with Freeze-Dried Spermatozoa," Biology of Reproduction 67(2), 409-415, (1 August 2002). https://doi.org/10.1095/biolreprod67.2.409
Received: 16 May 2001; Accepted: 1 February 2002; Published: 1 August 2002
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