Pregnancy after vitrification of pronuclear stage oocytes biopsied for polar body aneuploidy screening.

This paper reports the case of a patient undergoing IVF and polar body analysis for aneuploidy screening because of advanced maternal age. A total of nine fertilized oocytes were obtained from the patient's third treatment cycle. Three supernumerary pronuclear stage oocytes were identified as suitable after aneuploidy screening for five chromosomes and vitrified. Because the fresh cycle did not result in a pregnancy, vitrified pronuclear stage oocytes were warmed for transfer in an artificial cycle. All three zygotes survived warming and were transferred to the patient's uterus 2 days later as 4-, 5- and 6-cell stage embryos. After 2 weeks, a positive pregnancy test indicated successful implantation. The gestation is developing normally and is presently (end of September 2007) in the 22nd week.

The outcome of intracytoplasmic sperm injection in patients with retrograde ejaculation.

The objective of this retrospective clinical study was to assess the benefit of assisted fertilization in cases of anejaculatory infertility due to retrograde ejaculation. We report the outcome of intracytoplasmic sperm injection (ICSI) treatment. In 16 couples in which the men suffered from retrograde ejaculation. We performed 35 cycles of ICSI with spermatozoa retrieved from post-ejaculatory urine. The patients had been instructed to alkalinize the urine by ingesting sodium bicarbonate before the procedure. The fertilization rate averaged 51.2%. Seven clinical pregnancies were achieved. Three spontaneous first trimester abortions occurred, but three live offspring were delivered and one pregnancy is ongoing. In conclusion, the use of ICSI may be feasible for patients with retrograde ejaculation who are resistant to medical treatment and whose sperm quality is so low or unpredictable that intrauterine insemination or conventional methods of in-vitro fertilization are not possible.

High-frequency electric field trapping of individual human spermatozoa.

We present a new touch-free technique for trapping, positioning and selecting human spermatozoa. This can be done in free solution (culture medium) by high-frequency electric fields. Ultramicroelectrodes fabricated by photo- and electron-beam lithography on quartz and glass substrates were used to create field cages or long field channels. If the conductivity of the external salt solution is higher than the average value of sperm cell conductivity, negative polarization and negative dielectrophoresis occur. As a result, the induced cell polarization leads to forces repelling spermatozoa from the electrodes towards the field minimum. Using four planar electrodes a field funnel can be formed in which an individual spermatozoon is retarded while swimming. The same can be done more effectively in three-dimensional cages created by an octopole electrode system. In these systems, rapidly swimming spermatozoa could be trapped for several seconds but some spermatozoa stop moving if exposed to field strengths of more than 500 V/cm at frequencies in the MHz range. However, in stripwise and interdigitated electrodes, rapidly swimming sperm cells could be very well positioned in front of a break-electrode by a combination of electric field trapping and field induced laminar fluid streaming. This technique can be applied to bring individual spermatozoa to a defined position for characterization followed by sampling with capillaries.

Reduced incidence of ovarian hyperstimulation syndrome by prophylactic infusion of hydroxyaethyl starch solution in an in-vitro fertilization programme.

Prophylactic infusion of human serum albumin can reduce or mitigate severe ovarian hyperstimulation syndrome (OHSS) in patients at high risk. Recently, concern has been expressed in the lay press regarding the potential viral transmissions with blood constituents. Hence, we looked for a safe non-biological substitute with comparable physical properties in order to cope with this concern. One hundred patients of our in-vitro fertilization (IVF) programme with oestradiol serum concentrations > or = 11010 pmol/l on the day of human chorionic gonadotrophin injection and/or > or = 20 oocytes retrieved and/or previous severe OHSS were infused with 1000 ml 6% hydroxyaethyl starch solution at the time of oocyte collection and 500 ml 48 h later. A total of 82 IVF patients at risk without prophylactic infusions during the preceding years served as controls. Both groups were identical according to patient's age, body mass index, androgen concentrations, peak oestradiol concentrations, number of retrieved oocytes, fertilization and pregnancy rates. There were seven cases of severe OHSS in untreated patients and two cases in the treatment group (P = 0.08). In moderate OHSS a significant difference became obvious with only ten cases in the treatment group and 32 cases in the control group (P < 0.00001). Hydroxyaethyl starch solution seems to be an effective and economic alternative in reducing severe and moderate OHSS during IVF treatment.

Pregnancy after intracytoplasmic sperm injection of spermatozoa extracted from frozen-thawed testicular biopsy.

The case report illustrates the successful application of a new method of sperm extraction from frozen-thawed testicular biopsy specimen within an established programme of intracytoplasmic sperm injection.

Long-term follow-up in 206 infertility patients with polycystic ovarian syndrome after laparoscopic electrocautery of the ovarian surface.

In recent publications we have demonstrated that laparoscopic electrocautery of the ovarian surface (LEOS) is an effective method to reduce serum androgen concentrations, normalizing ovarian cycle length and the ovarian reaction to hormonal stimulation therapy in anovulatory patients with polycystic ovarian disease (PCOD). In this paper we show that these effects are not only temporary. Data from 206 patients undergoing LEOS and monitored for up to 72 months after surgery are evaluated; 145 patients achieved a total of 211 conceptions, giving a pregnancy rate of 70%, with a maximum of four conceptions in one patient. There were 38 early miscarriages (18%) and two late pregnancy losses as well as three ectopic implantations.

Patterns of serum-luteinizing hormone surges in stimulated cycles in relation to injections of human chorionic gonadotropin.

Endogenous-luteinizing hormone (LH) surges may complicate the management of in vitro fertilization cycles. To investigate the effects of LH surges after hormonal stimulation 53 IVF cycles were analyzed by assessing LH levels three times daily until egg collection. In 43% the LH rise started before the planned exogenous trigger for ovulation was given, in 11% the rise occurred simultaneously with and in 45% after the injection of human chorionic gonadotropin. Three main patterns of serum LH surges were identified: (A) low-LH tonus with straight increase to maximum; (B) low tonus with elevation before straight increase; (C) high tonus with large variations but no prominant peak. These patterns were not related to the follicular estradiol increase, luteal steroid concentrations or resulting pregnancy rates.

Evaluation of endocrine parameters and semen samples: pregnancies following conservative infertility treatment versus IVF therapy.

IVF techniques have recently attracted interest as a possible treatment of male subfertility. In order to clarify if semen profiles of men fathering a child after IVF differ significantly from those patients initiating a pregnancy after timed intercourse or AIH, we retrospectively compared sperm parameters in 67 males whose female partners were treated by IVF with those of 102 couples treated conservatively. In all male partners stimulation tests for FSH and LH, TSH and prolactin (PRL) had been performed as well as measurement of testosterone (T) levels. No significant differences were revealed for mean values of any of the classical semen parameters: volume, concentration, vitality, motility, normal morphology and results of the hypoosmotic swelling test between groups for men initiating a conception. Endocrine evaluation of the male patients detected no major hormonal imbalances associated with specific subnormal sperm parameters although regression analyses demonstrated relationships between semen volume and TSH as well as T, and between sperm concentration and PRL as well as FSH levels. We conclude that IVF techniques are not superior to cycle optimization with timed insemination in the treatment of andrological subfertility.

[Effect of various factors on in vitro fertilization (IVF) and embryo transfer in the human in clomiphene/hMG/hCG stimulated cycles]. / Einfluss verschiedener Faktoren auf die In-vitro-Fertilisation (IVF) und den Embryotransfer beim Menschen in Clomiphen/hMG/hCG-stimulierten Zyklen.

In the present study we tried to analyse data obtained on the occasion of an in vitro fertilization programme at the department of obstetrics and gynecology of the University of Kiel, to extract some of the important factors, because their positive or negative influence on the success of extracorporal insemination, embryo transfer and implantation has not been clearely defined till now and to value their concern. Between July 1982 and February 1983 105 sterile patients with ovulatory menstrual cycles have been treated by the technique of in vitro fertilization and embryo transfer, successively. Follicular growth was stimulated by 150 mg clomiphen daily from 3rd to 7th day and application of 225 IU hMG on 6th, 8th and 10th day. Finally ovulation was timed 36 hours before follicular puncture by 5000 IU hCG. In 24 per cent endogenous LH peak could be observed. In 86 per cent of the 80 patients exactly timed one oocyte was to be found minimally. In 68 per cent of these patients after all an embryo transfer was done (24 times one embryo, 13 times two, 9 times three and once four embryos). 89% of the embryos have been transfered as 4-or 8-cell stage, only 11 per cent as two cell stage. Transfer took place 48 to 50 hours following insemination. There was no correlation between the maturity classified morphologically of the oocytes and the diameter of their follicles. The speed of development of the embryos was not correlated with the size of their follicles. too. In 60-70 per cent of all cases, in which an embryo transfer was possible, husbands had normal spermiogram. The percentage of unfertilised oocytes following insemination using non-optimal sperma was increased in all types of cycles, additionally there was a reduction of the developing speed of resulting embryos. Doubtless two the four pregnancies resulted after insemination using sperma of less quality. The conjugation rate is said to be not correlated directly with the exactness of timing of the application of hCG with reference to follicular maturity, but in all pregnancies hCG was injected simultaneously with the peak of estradiol.

Prolactin concentrations in follicular fluid following ovarian hyperstimulation for in vitro fertilization.

Prolactin (PRL) and sex steroid concentrations were measured in follicular fluids of women treated either with (1) clomiphene/hCG or with (2) clomiphene + hMG/hCG. Method 1 of ovarian stimulation resulted in lower follicular PRL and higher oestradiol-17 beta (E2) and progesterone (P) concentrations than method 2. There was no difference in the PRL and sex steroid concentrations of follicles with fertilized and of those yielding unfertilized ova, but in both stimulation types, follicles from which no oocytes were obtained had high PRL and low E2 and P levels. Significant positive correlations were evident for PRL and T and E2 and P, respectively, while PRL and P were negatively correlated.

[Pregnancy and birth of monozygous female twins following in vitro fertilization and embryo transfer]. / Schwangerschaft und Geburt monozygoter weiblicher Zwillinge nach In-vitro-Fertilisation und Embryotransfer (IVF-ET).

The rate of identical twins after in vitro fertilisation and embryo replacement is higher than after physiological conception. The present paper reports on the first delivery of twins (identical twins) after in vitro fertilisation and embryo replacement. The in vitro fertilisation and embryo replacement of one 4-cell stage resulted in a twin pregnancy. The difficult pregnancy, early delivery and intense perinatal care of the children deserves special consideration. In addition the paper gives a survey on the present number of deliveries after in vitro fertilisation and embryo replacement given at the 3rd World Congress on In Vitro Fertilisation and Embryo Transfer at Helsinki in May 1984.

[Is the sperm bacteriology important for the results of in vitro fertilization?]. / Haben bakteriologische Befunde des Spermas für die in vitro-Fertilisation Bedeutung?

A total of 140 ejaculates from patients taking part in the in vitro fertilization program were examined. In 60 ejaculates (42.9%) bacteria concentrations of greater than 10(5)/ml were found. 80 (57.1%) of the probes were negative or had bacteria concentrations less than 10(4)/ml. Microplasma was found in 30 of the examined ejaculates (21.4%) and the second most frequent group of microorganisms were enterococci found in 15 patients (10.7%). Anaerobic organisms in concentrations of more than 10(5)/ml were detected. After insemination of one of more pelviscopically obtained oocytes with sperm of the husband in 14 out of 22 cases (63.6%) with bacteria concentrations greater than 10(5)/ml the cleavage of at least one oocyte occurred leading to an embryo transfer rate of 63.6%. 14 of 31 ejaculates with negative bacteriological findings (45.2%) fertilized at least one of the inseminated oocytes. From the results obtained up to now any direct effect of microorganism contaminated sperm on the human in vitro fertilization system resulting in reduced fertilizations rates cannot be demonstrated. It can be said that there is no correlation between microbiological and cytological findings of ejaculates used for insemination of oocytes and the obtained fertilization, cleavage and embryo transfer rates in the human in vitro fertilization program here. But since the negative effect on implantation rates or embryonic development cannot be excluded detailed bacteriological examinations and probably antibiotic treatments may be recommended before admission of the in vitro fertilization program.

Monoclonal sperm antibodies: their potential for investigation of sperms as target of immunological contraception.

We generated 149 hybridoma cell lines secreting antibodies against human spermatozoal antigens of which antibodies from 136 hybridoma lines also reacted with seminal plasma constituents. The occurrence of common antigeneic determinants on spermatozoa and seminal plasma was further demonstrated by competitive inhibition ELISA tests. We found that seven hybridoma clones secreted antibodies reactive with spermatozoa but nonreactive with seminal plasma. Antibodies from 5 clones were sperm-agglutinating and from 15 clones sperm-immobilizing. Localization of sperm antigens reactive with the monoclonal antibodies was demonstrated by indirect immunoperoxidase staining. A synthetic decapeptide, earlier shown to be reactive with naturally occurring human iso- and autosperm antibodies, was shown to be reactive with the monoclonal antibody VII-5 in ELISA tests.

Minimal requirements for ejaculates used for in vitro fertilization.

Minimal requirements for ejaculates to be used for in vitro fertilization of human oocytes are defined. Experimental results obtained a total concentration of at least 5 X 10(6) spermatozoa per ml are required, with at least 30% of the sperm showing normal motility and normal morphology in the differential spermiocytogram. The total volume of ejaculate should not be less than 1 ml. A total of 140 ejaculates from patients taking part in the in vitro fertilization program were examined. In 60 ejaculates (42.9%) bacteria concentrations of greater than 10(5)/ml were found. Eighty (57.1%) of the probes were negative or had bacteria concentrations less than 10(4)/ml. Mycoplasma was found in 30 of the examined ejaculates (21.4%) and the second most frequent group of microorganisms were enterococci found in 15 patients (10.7%). Anaerobic organisms in concentrations of more than 10(5)/ml were not detected. After insemination of one or more pelviscopically obtained oocytes with sperm of the husband in 14 out of 22 cases (63.6%) with bacteria concentrations greater than 10(5)/ml the cleavage of at least one oocyte occurred leading to an embryo-transfer rate of 63.6%. In 14 of 31 ejaculates with negative bacteriological findings (45.2%) at least one of the inseminated oocytes was fertilized. Direct effect of microorganism contaminated sperm on the human in vitro fertilization system resulting in reduced fertilization rates cannot be demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS)

Induction of high titer mouse--antihuman spermatozoal antibodies by liposome incorporation of spermatozoal membrane antigens.

This is the first report of induction of high titer mouse-antihuman spermatozoal antibodies using a spermatozoal antigen incorporated in liposomes. The peptide antigens that were incorporated into pure phosphatidylcholine liposomes are known to react with the naturally occurring antibodies in sera of sterile patients. Electron microscopy and the complement-dependent glucose release tests indicated that the peptides were entrapped in the aqueous phase between the lipid bilayers in the liposome vehicles. This is in contrast to most other liposomal antigens that are expressed on the outer lipid surface. In view of the nontoxic nature of the liposomes these results could be of some interest in immunological fertility regulation trials with appropriate antigens in an allogeneic system.