Extraction of testicular sperm from previously cryopreserved tissue in couples with or without transport of oocytes and testicular tissue.

OBJECTIVE: To evaluate results of IVF and intracytoplasmic sperm injection (ICSI) with extraction of sperm from frozen-thawed testicular tissue. DESIGN: Retrospective follow-up study. SETTING: Fertility center. PATIENT(S): Thirty-five couples with transport of testicular tissue from a transport clinic and 125 local couples. INTERVENTION(S): Extraction of testicular sperm by maceration and enzymatic digestion from frozen-thawed testicular tissue before ICSI. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate (PR) and implantation rate in couples with obstructive or nonobstructive azoospermia, motile or immotile sperm, and differing male serum FSH values. RESULT(S): The clinical PR per ET and implantation rate per embryo in couples with transport of testicular tissue were 40% and 18%, respectively, in cases of obstructive azoospermia and 37% and 26%, respectively, in cases of nonobstructive azoospermia. In the local couples, these rates were 42% and 19%, respectively, in cases of obstructive azoospermia and 18% and 10%, respectively, in cases of nonobstructive azoospermia. The implantation rates for ICSI were 26% with motile sperm and 11% with immotile sperm in the transport group and 16% and 8%, respectively, in the local group. Male serum FSH level did not clearly correlate with implantation rate. CONCLUSION(S): Clinical PR and implantation rate are not affected by transport of testicular tissue but are significantly affected by nonobstructive azoospermia and the use of immotile sperm. No major increase in chromosomal aberration or congenital malformation was noted in the offspring of this limited group.

Chromosomal aberrations in couples undergoing intracytoplasmic sperm injection: influence on implantation and ongoing pregnancy rates.

OBJECTIVE: To determine the incidence of chromosomal aberrations in couples undergoing intracytoplasmic sperm injection (ICSI) and their influence on subsequent implantation and ongoing pregnancy rates. DESIGN: Prospective study. SETTING: Fertility center. PATIENT(S): Candidates for ICSI. INTERVENTION(S): Chromosomes were trypsin-banded in 2,280 patients. In all cases, 10 metaphases were karyotyped. Sex chromosome analysis was performed in 10 additional metaphases. When apparent chromosomal aberrations were detected, 100 metaphases were analyzed. MAIN OUTCOME MEASURE(S): Implantation and ongoing pregnancy rates in couples with a chromosomal disorder. RESULTS: A chromosomal abnormality was demonstrated in 7.2% of all couples. Among the male partners, 4.48% had aberrations. Autosomal aberrations were present in 2.96%, and numerical or structural sex chromosome abnormalities were found in 1.52%. Among the female partners, numerical or structural abnormalities were documented in 9.79%. Only 2.32% of the female partners had autosomal structural abnormalities. Numerical or structural anomalies involving sex chromosomes were found in 7.47%. Implantation rates of 9.4% and 16.3% per embryo were observed in female partners with sex chromosome mosaicism and autosomal aberrations, respectively. In male partners, the respective rates were 3.8% and 23.1%. CONCLUSION(S): The incidence of chromosomal disorders in couples seeking ICSI treatment is considerable, especially minor mosaicism (<10%) of sex chromosomes in the female partners. Preliminary data indicate a low implantation rate in couples with minor mosaicism of sex chromosomes.

Blastocyst transfer in day-5 embryo transfer depends primarily on the number of oocytes retrieved and not on age.

OBJECTIVE: To analyze the effects of patient age and treatment cycle number on the occurrence of blastocyst transfer and subsequent implantation. DESIGN: Prospective study. SETTING: Department of endocrinology and reproduction. PATIENT(S): All 1,099 women had day-5 transfers after IVF or intracytoplasmic sperm injection treatment. INTERVENTION(S): All patients were checked for embryo development in vitro in consecutive day-5 transfer cycles. Two blastocysts or three lesser-developed embryos were transferred. MAIN OUTCOME MEASURE(S): Blastocyst formation rate or clinical pregnancy/implantation rate. RESULT(S): Of 929 patients in the first cycle, 545 (59%) had at least one blastocyst available for ET. Among 151 patients with a blastocyst in cycle 1, 77 developed one or more blastocysts in cycle 2 (51%). Fifty of 143 patients without a blastocyst in cycle 1 had at least one blastocyst in cycle 2 (35%). After subdivision of all day-5 ETs according to the first four cycles, the following implantation rates per embryo were found for ET with one or more blastocysts: cycle 1 (n = 545), 23%; cycle 2 (n = 264), 23%; cycle 3 (n = 110), 14%; and cycle 4 (n = 27), 12%, and with noncavitating embryos, respectively: (n = 384) 6%, (n = 193) 6%, (n = 94) 2%, and (n = 35) 3%. The negative correlation of the age of the woman on blastulation depended primarily on the number of oocytes retrieved. CONCLUSION(S): The blastocyst implantation rate decreased after cycle 2. Biologic ovarian age, rather than chronologic age, determines the frequency of blastocyst transfer or pregnancy rate.

A prospective, randomized study of embryo transfer results after 3 or 5 days of embryo culture in in vitro fertilization.

OBJECTIVES: To compare the implantation rate of embryos after 3 and 5 days of IVF culture. DESIGN: Prospective randomization of ET depending on the weekday of ovum pick-up (OPU). SETTING: University Department of Endocrinology and Reproduction. PATIENTS: All women entering an outclinic IVF program. INTERVENTIONS: Two hundred thirty-three ETs performed on day 3 after OPU and 410 performed on day 5 were analyzed. When blastocysts with a clear inner cell mass were available, a maximum of two were replaced. RESULTS: On day 3 after OPU, 60 pregnancies per 233 ET (26%) and on day 5, 102 pregnancies per 410 ET (25%) were induced. The average implantation rate per embryo was 13% and 12%, respectively. After subdivision according to embryo morphology, pregnancy rate per ET (n = 59) and implantation rate per embryo on day 3 with exclusively unfragmented embryos were 32% and 18%, respectively, not significantly different from ET (n = 73) exclusively with embryos containing > 0% and < 20% fragments: 27% and 12%. After transfer on day 5, when one or more cavitating embryos were available (n = 227), pregnancy and implantation rates were 40% and 23%, statistically different from ET on day 3. On day 5, ET exclusively with morula stages showing signs of starting blastulation (n = 26), pregnancy rate and implantation rate were 12% and 11%, respectively, from ET (n = 157) with embryos not reaching the latter stage: 6% and 3%. CONCLUSIONS: Overall ET results after 3 and 5 days are comparable. After 5 days of culture, one to two embryos can be replaced with an average implantation rate of > 23% per embryo, minimizing the incidence of triplets.

A randomized study of transcervical intrafallopian transfer of pronucleate embryos controlled by ultrasound versus intrauterine transfer of four- to eight-cell embryos.

OBJECTIVE: To establish the value of transcervical intrafallopian transfer of zygotes and the accuracy of fallopian cannulation. DESIGN: A prospective randomized study, comparing ultrasonically controlled transcervical intrafallopian transfer of zygotes with intrauterine transfer of cleaved embryos. SETTING: Department of IVF of the Rotterdam Academic Hospital. PATIENTS: One hundred forty-five patients with patent tubes entered the IVF program. MAIN OUTCOME MEASURE: Pregnancy rates in both groups and ultrasound (US) assessment during fallopian cannulation and ET. RESULTS: Transvaginal cannulation of the tube appears not to be sufficiently accurate when performed without US guidance. Catheter damage occurred in many cases. The previously reported superior implantation rate after intrafallopian transfer in comparison with intrauterine transfer could not be confirmed. CONCLUSIONS: Intrauterine transfer of cleaved embryos remains the method of choice in IVF.

Vaginal disinfection with povidon iodine and the outcome of in-vitro fertilization.

In a prospective randomized study, the influence was assessed of vaginal disinfection with a 1% solution of povidon iodine (Betadine), before performing a transvaginal ultrasound-guided oocyte retrieval, on fertilization, cleavage rate and pregnancy rate. The outcome of 334 oocyte retrievals was studied. In 160 cases, Betadine was used and in the remaining 174 cases, normal saline was used. No differences in the fertilization and cleavage rates were found (fertilization 45.5% versus 47.8%, cleavage 49.8% versus 52.1% in the Betadine and normal saline groups respectively). However the pregnancy rate was significantly higher in the normal saline group (17.2% versus 30.3% clinical pregnancies per embryo transfer). No increase in infection risk occurred in the saline group.

Transcervical intrafallopian transfer of zygotes.

Reportedly, gamete intrafallopian transfer and zygote intrafallopian transfer are successful methods in assisted conception. This pilot study describes the experiences and results of a recently developed technique of vaginal transcervical intrafallopian transfer. In a group of 38 women with unexplained infertility, oocytes were retrieved. In 25 patients, pronucleate embryos were transferred to the fallopian tubes. A positive pregnancy test was reported in 8 cases. Considerations pertaining to this method and technical implications are discussed.

Uterine and ovarian flow velocity waveforms in the normal menstrual cycle: a transvaginal Doppler study.

A combined transvaginal 2D real-time and pulsed Doppler method was used for recording flow velocity waveforms in the uterine and ovarian arteries from 16 healthy women during the follicular and luteal phase of the normal menstrual cycle. Continuous forward end-diastolic flow velocities were documented in 74% of the ovarian artery and 96.5% of the uterine artery flow velocity waveforms. Comparison of the pulsatility index from the left and right ovarian artery revealed a significantly lower pulsatility index on the side of the ovary bearing the developing corpus luteum, suggesting reduced down-stream impedance or increased blood flow. The pulsatility index from the uterine artery only seems to be marginally involved in the observed impedance changes during the luteal phase of the menstrual cycle.

The influence of the interval between in vitro fertilization and embryo transfer and some other variables on treatment outcome.

This prospectively randomized study compares the outcome of 434 ETs 48 to 52 hours after insemination (day 2) and 324 ETs 72 to 76 hours after insemination (day 3). The influence of the interval between insemination and ET was assessed, as well as that of the number of embryos transferred, embryo quality, and the presence of supernumerary embryos. The mean number of embryos transferred after 2 and 3 days was equal (2.9 embryos/ET). The pregnancy rates per ET were not significantly different (21.9% versus 23.5%), but a higher percentage of viable pregnancies was observed after ET on day 3 (88.2% against 69.5% following ET on day 2). Treatment outcome was positively correlated with the number of embryos transferred and the presence of supernumerary embryos.

The value and role of plasma 17 beta-oestradiol measurements during ovarian hyperstimulation for in-vitro fertilization.

Follicular aspiration was performed in 464 hyperstimulated IVF treatment cycles in patients with severe tubal damage as the sole cause of their infertility. In 413 cycles, one to four embryos could be replaced, resulting in 102 clinical pregnancies. In 458 treatment cycles, data on plasma E2 levels were available on days -3 and -2, in 322 cycles also on days -1 and 0, day 0 being the day of follicular puncture. Although the distribution of cycles leading to clinical pregnancy within the 5-95th centile for plasma E2 levels differed from that observed outside this range, these differences were of no statistical significance. These results indicate that IVF pregnancies occur in the presence of a wide range of E2 levels, during the 3-day period preceding follicular aspiration. The importance of plasma E2 measurements for treatment policy must, therefore, be reconsidered.

Factors influencing the outcome of successive IVF treatment cycles in attaining a follicular puncture.

An analysis has been made on data from 576 cycles in 300 patients regarding the chances of various superovulation protocols attaining a puncture in successive cycles. The cumulative proportion of patients with at least one successful stimulation increased from 77% at the first attempt, to 98% at the fifth attempt. Of the 576 stimulations, 440 (76%) resulted in a puncture. Age, the number of ovaries and the reaction to superovulation induction contribute to the outcome of a cycle, the chances being lowest for older individuals with one ovary and an inadequate reaction in the first treatment cycle. Changing the treatment protocol did not improve the outcome in subsequent gonadotrophin-induced cycles, even if combined with clomiphene citrate. Reasons for cancellation did not tend to recur, although the incidence of dominance and premature stimulation was significantly higher in patients aged greater than 35 years. Age-dependent chances of success were calculated.

The factor VIII ratio in normal and pathological pregnancies.

The factor VIII ratio was measured in 102 pregnant women (43 uncomplicated pregnancies, 19 with pre-eclampsia without fetal growth retardation, 8 pre-eclampsia complicated by fetal growth retardation, 12 with only fetal growth retardation and 20 with gestational diabetes). The pathological pregnancies showed a statistically significant elevation of the factor VIII ratio compared to the normal pregnancies. The group of pre-eclampsia complicated by fetal growth retardation showed the most marked increase of the factor VIII ratio. The results are discussed in relation to the hypothesis that the elevated factor VIII ratio is mainly due to the release of factor VIII antigen by various forms of vascular stress.