Prospective, auto-controlled study on reinsemination of failed-fertilized oocytes by intracytoplasmic sperm injection

To evaluate the possible influence of late fertilization after standard IVF on the results of reinsemination of assumed failed-fertilized oocytes by microinjection and to examine the correlation between the effect of aging of (failed-fertilized) oocytes and the ability of these oocytes to become fertilized.

Trial 1: Group 1 (injected-day 1), 93 failed-fertilized oocytes injected 1 day after ovum pick-up; group 2 (control), 82 failed-fertilized oocytes with no microinjection performed. Trial 2: Group 1 (ICSI-day 1), 40 failed-fertilized oocytes injected 1 day after ovum pick-up; group-2 (ICSI-day 2), 40 failed-fertilized oocytes injected 2 days after ovum pick-up. In addition, 35 two- to eight-cell stage embryos, obtained after ICSI of IVF failed-fertilized oocytes, were fixed for cytogenetic analysis.

Normal and abnormal fertilization and embryo development.

Trial 1: 53% normal (2 pronuclear [PN]) and 25% abnormal (> or = 3PN) fertilization rates were obtained in group 1 (injected-day 1), and 71% of the 2PN and 74% of the > or = 3PN oocytes cleaved with < 50% fragmentation. No pronuclear (> or = 2PN) development occurred in the control group. Trial 2: 45% and 8% normal and 25% and 40% abnormal fertilization rates were obtained, respectively, after ICSI of 1-day-old and 2-day-old failed-fertilized oocytes. Two days after microinjection, 67% and 67% of the 2PN and 80% and 44% of the > or = 3PN oocytes cleaved with < 50% fragmentation in group ICSI-day 1 and in group ICSI-day 2, respectively.

Late fertilization after initial in vitro insemination does not play a role in the high fertilization rate obtained after reinsemination of assumed failed-fertilized oocytes by ICSI. Normal (2PN) fertilization rate, however, decreases strongly and the abnormal (> or = 3PN) fertilization rate increases with oocyte aging and derived embryos seem to have a high incidence of cytogenetic abnormalities.