Birth of normal calves after intracytoplasmic sperm injection of bovine oocytes: a methodological approach.

摘要: Intracytoplasmic sperm injection (ICSI) is advantageous when only very few spermatozoa are available for insemination. Bovine were injected individually into matured oocytes using a piezo electric actuator. Spermatozoa "immobilized", by scoring their tails immediately before injection, or "killed", repeated freezing and thawing. About 4 h after ICSI, the with two polar bodies (activated injection) selected treated 5 min 7% ethanol further culture. When examined 19-21 nearly 90% of fertilized normally (two pronuclei bodies) irrespective treatment (immobilization killing) prior to but subsequent preimplantation embryo development was much superior (cleavage 72%: blastocysts 20%) ICSI immobilized than killed 28%; 1%). Ethanol activation bovine improved cleavage (33% versus 72%) blastocyst (12% rates markedly (P < 0.05). Five normal calves born transplantation ten surrogate cows. These results show that piezo-ICSI spermatozoa, combined sperm-injected oocytes, an effective method produce offspring.