Avoiding multiple pregnancies in ART

The effectiveness of blastocyst culture and transfer in human in-vitro fertilization (IVF) was evaluated in a prospective randomized trial in patients having a moderate to good response to gonadotrophin stimulation. Embryos were transferred either on day 3 after culture to around the 8-cell stage in Ham's F-10 medium supplemented with fetal cord serum, or on day 5 after culture to the blastocyst stage in the sequential serum-free media G 1.2 and G 2.2. The pregnancy rates after transfer on day 3 or day 5 were equivalent, 66 and 71% respectively; however, significantly more embryos were transferred on day 3 (3.7) than on day 5 (2.2). The number of blastocysts transferred did not affect the implantation rate, and pregnancy rates when either two or three blastocysts were transferred were 68 and 87% respectively. The implantation rate of the blastocysts (50.5% fetal heart beat) was significantly higher compared to the cleavage stage embryos transferred on day 3 (30.1%). The percentage of blastocyst development was not affected by the number of 2-pronuclear embryos, or by maternal age. Irrespective of the number of blastocysts formed, pregnancy rates were similar. Furthermore, the pregnancy rate following blastocyst transfer in patients with 10 or more follicles at the time of human chorionic gonadotrophin administration was not affected by patient age. More than 60% of patients having blastocyst culture and transfer had supernumerary embryos for cryopreservation. The establishment of a pregnancy following thaw and transfer confirmed the viability of cryopreserved blastocysts cultured in the absence of serum or co-culture. The ability to transfer just two blastocysts while maintaining high pregnancy rates will therefore help to eliminate high order multiple gestations and improve the overall efficiency of human IVF.

In most in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) programmes approximately one ongoing pregnancy in three is multiple. The need to characterize embryos with optimal implantation potential is obvious. We retrospectively examined all of 23 double transfers resulting in ongoing twins, occurring between January 1, 1996 and May 19, 1997. Characteristics of these top quality embryos were absence of multinucleated blastomeres, four or five blastomeres on day 2, seven or more cells on day 3, and 2 embryos, 11/31 (35%) were multiple. We applied our top quality criteria to the 221 double transfers: 106 transfers with two top embryos resulted in 65 (63%) ongoing pregnancies with 37 (57%) twins, 65 transfers with one top embryo in 38 (58%) ongoing pregnancies with eight (21%) twins. In the group without top embryos, 12/52 (23%) ongoing singletons occurred, with no twins. The corresponding ongoing implantation rates were 49, 35 and 12%. This analysis suggests that single embryo transfer with an acceptable pregnancy rate might be considered if a top quality embryo is available.

This retrospective study was undertaken to determine whether further developmental progression of two-pronucleated (2PN) zygotes can be predicted by a single, non-invasive examination of pronuclei, with the use of criteria based on the number and distribution of nucleolar precursor bodies in each pronucleus. The normal range of pronuclear variability was defined by analysis of zygotes giving rise to embryos transferred in 100%-implantation cycles (pattern 0). Morphological patterns differing from pattern 0 were classified as patterns 1-5. The frequency of developmental arrest of pattern 0 zygotes was only 8.5% as compared with 31.6, 21.9, 30.0, 20.5 and 24. 1% for patterns 1-5 respectively. Relationships of pronuclear patterns with blastomere multinucleation and cleaving embryo morphology were also noted. Clinical pregnancy was achieved in 22 of 44 (50%) treatment cycles in which at least one pattern 0 embryo was transferred, but only in two of 23 (9%) cycles in which only pattern 1-5 embryos were transferred. These data present new evaluation criteria which can be used to predict the developmental fate of human embryos as early as the pronuclear stage, without requiring repeated observations or an exact timing of pronuclear zygote inspection. Further prospective study is needed for clinical validation of these criteria.