Impact of three-dimensionally measured endometrial volume on the pregnancy rate after intrauterine insemination.

OBJECTIVE: The success of artificial reproductive techniques not only depends on the quality of oocytes and spermatozoa but also on the receptivity of the endometrium. The aim of this study was to assess the role of endometrial volume measurement by three-dimensional ultrasound in predicting the pregnancy rate (PR) in women receiving controlled ovarian hyperstimulation followed by intrauterine insemination. MATERIAL AND METHODS: 104 patients having intrauterine insemination (IUI) were included in this prospective trial. Ovarian hyperstimulation was performed with gonadotropins in 73 % of patients and with clomiphene citrate (CC) in 27 % of patients. Endometrial thickness, pattern and three-dimensional volume were measured immediately before insemination. RESULTS: In 104 IUI cycles a total of 14 clinical pregnancies were recorded (PR=13.5 %). The endometrial volume was 3.5 ml and was not significantly different in pregnant (4.0 +/- 1.5 ml) from non-pregnant women (3.4 +/- 1.9 ml). In the subgroup of women with an endometrial volume > or = 2 ml and trilaminar endometrium the pregnancy rate was 22 %, significantly higher than that in women without these two criteria (PR 6 %, p < 0.05). The negative predictive value of an endometrial volume < 2 ml for a clinical pregnancy after IUI was 96 %. Endometrial volume and thickness were significantly higher after ovarian hyperstimulation with gonadotropins (3.7 ml and 11.0 mm) than with CC (2.8 ml and 9.5 mm; p < 0.05). CONCLUSIONS: An endometrial volume < 2 ml at the day of insemination is associated with a poor likelihood of pregnancy. Endometrial volume measured by 3D ultrasound is a new objective parameter to predict endometrial receptivity.

[First experiences with human blastocyst culture after IVF/ICSI under the conditions of the German embryo protection law]. / Erste Erfahrungen mit der Blastozystenkultur nach IVF oder ICSI unter den Bedingungen des Deutschen Embryonenschutzgesetzes.

OBJECTIVE: Due to the improvements in human embryo culture in the recent years, it is now possible to transfer embryos five days after oocyte retrieval and IVF or ICSI at the blastocyst stage with favorable implantation rates. In Germany it is illegal to cultivate more than 3 embryos, therefore the selection has to be done at the pronuclear stage. There we report our experiences of human blastocyst culture in a routine IVF/ICSI programme under the conditions of the German Embryo Protection Law. MATERIALS AND METHODS: The data of 100 couples undergoing the IVF-ICSI programme at the University Clinic of Würzburg were analysed prospectively. 14-18 hours after insemination or micro-injection two or three zygotes with the best pronuclear development were selected for further cultivation. Fertilized oocytes were cultured in sequential media and were then transferred into the uterus 5 days after oocyte recovery. The blastocysts were graded from 1-8. RESULTS: In 100 cycles a total of 859 oocytes were collected, of whom 663 were fertilized and reached the pronuclear stage (median fertilization rate 88.9 %). 251 zygotes were selected at the PN stage. 51 % of the selected zygotes achieved the blastocyst stage after 5 days (grade 1-5), 28 % were morulae (grade 6-7) and 21 % of the embryos arrested in their development (grade 8). The clinical pregnancy rate was 26 %. Women who conceived had a significant better development of blastocysts on day 5 (grade 4 versus grade 6, P < 0.01) than those not achieving pregnancy. CONCLUSIONS: In summary, under the current legal conditions in Germany, blastocyst culture cannot improve pregnancy rates as the rate of arrested embryos of over 20 % limits the chances of implantation.

The use of a detailed zygote score after IVF/ICSI to obtain good quality blastocysts: the German experience.

BACKGROUND: Due to improvements in embryo culture, it is now possible to transfer embryos 5 days after oocyte retrieval and IVF/ICSI at the blastocyst stage, giving a better synchronization with the female reproductive tract. In Germany it is illegal to culture more than three embryos. Therefore, there is need for a sufficient selection at the pronuclear (PN) stage to select the best zygotes and exclude those of poor quality. METHODS: A prospective trial was conducted in 168 IVF and ICSI cycles including the size, number and alignment of pronuclei and nucleoli, cytoplasmic halo effect, the presence of vacuoles and granularity of ooplasm. Based on the above criteria, the best zygotes were selected (score <15) for embryo transfer on day 5. Blastocysts were classified in eight grades based on the cleavage speed. RESULTS: A total of 1450 oocytes were collected, of which 1119 reached the pronuclear stage. Of the zygotes (n = 424) selected at the PN stage, 46% achieved the blastocyst stage after 5 days (grade 1-5), 26% the morula stage (grade 6-7) and 28% were arrested (grade 8). The mean zygote score showed a significant positive correlation with the mean blastocyst quality in ICSI, but not in IVF cycles. A cut-off of 15 was calculated for ICSI cycles giving the best discrimination with blastocyst grades (6 versus 7) and number of arrested embryos (23 versus 45%) below and above this cut-off. A total of 33 clinical pregnancies was achieved (20%). Women conceiving had a significantly better mean blastocyst development than those not conceiving. Strong cytoplasmic vacuolization and an extreme or no halo effect had a negative effect on blastocyst development. CONCLUSIONS: The data show that PN stage morphology is related to blastocyst development, but the rate of arrested embryos of almost 30% limits the chance of conception under the conditions of the German Embryo Protection Law.

Semen sample collection in medium enhances the implantation rate following ICSI in patients with severe oligoasthenoteratozoospermia.

Ejaculation in medium increases the proportion of antibody-free spermatozoa in semen samples containing anti-sperm antibodies and thereby enhances the fertilization rate in vitro. The aim of this study was to investigate whether this technique is also beneficial in semen samples with severe oligoasthenoteratozoospermia (OAT) where bacteria and detritus are often present. A prospective randomized controlled trial was carried out to study the results of sperm preparation and fertilization and pregnancy rates after intracytoplasmic sperm injection (ICSI) for OAT. Of the 114 couples (one cycle per couple) studied between 1998 and 2000, 55 men were randomized to have semen collection into sterile dry pots (group A) and the remaining 59 had samples collected into 20 ml HEPES buffered Ham's F-10 medium with 10% human serum albumin (group B). In group B the ejaculates were incubated for 30 min and mixed gently. The samples were then processed by mild centrifugation and washing followed by a mini-swim-up technique. The ejaculates in group A were prepared by the swim-up procedure only. The overall fertilization rate was 71.8% and was similar in groups A (fertilization rate = 66.7%) and B (fertilization rate = 64.3%). In group A, 10/55 clinical pregnancies were recorded (pregnancy rate 18%), with an implantation rate (IR) of 6.9% per embryo. In group B, 16 of 59 patients conceived leading to significantly higher implantation (9.9%, P < 0.001) and clinical pregnancy rates (27%, P < 0.001). It is postulated that the addition of medium before liquefaction could inhibit the binding of bacteria and detritus to the sperm surface and may diminish DNA damage caused by reactive oxygen species, leading to improved efficiency of fertilization. The results demonstrate that the addition of HEPES buffered Ham's F-10 medium to sample collection pots significantly improves the pregnancy rate after ICSI in patients with severe OAT.