Evaluation of a TEST-yolk sperm capacitation system for use in bovine in vitro fertilization.

Abstract

Bovine sperm acquire the ability to penetrate zona-free hamster oocytes (capacitation) after incubation in TEST-yolk buffer. Our objective was to determine whether such sperm could penetrate zona-intact bovine oocytes in vitro. Bovine cumulus enclosed oocytes from 2- to 5-mm follicles were incubated in maturation medium for 24 h at 37 degrees C. Ejaculated bovine semen was diluted 1: 10 in TEST-yolk buffer, cooled to 4 degrees C, and stored for 8 h to induce capacitation. Sperm were then washed thrice in pH 7.6, .15 M NaCl containing .1% bovine serum albumin V (37 degrees C) and diluted to 2 x 10(6) sperm/ml in fertilization medium. Droplets of fertilization medium containing capacitated sperm, killed sperm, or no sperm were made under paraffin oil. Oocytes (matured 24 h) were added and cocultured with sperm for 8 h and then transferred to fresh fertilization medium for 40 h. After 24 h, 53% of the oocytes had matured (metaphase II). The fertilization rate of the metaphase II oocytes (203) with TEST-yolk capacitated sperm was 87%, whereas the parthenogenetic controls were 2 and 0%, respectively. Therefore, TEST-yolk buffer can be used to capacitate bull sperm for in vitro fertilization.

• Authors: A Ijaz, A G Hunter
• Journal: Journal of dairy science (J Dairy Sci) Vol. 75 Issue 2 Pg. 394-8 (Feb 1992) ISSN: 0022-0302 [Print] United States
• PMID: 1560133 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Buffers
• Culture Media

Topics

• Animals
• Buffers
• Cattle (physiology)
• Cells, Cultured
• Culture Media
• Female
• Fertilization in Vitro
• Male
• Meiosis
• Metaphase
• Oocytes (growth & development, physiology)
• Sperm Capacitation
• Sperm-Ovum Interactions