Chromosome studies of early human embryos: proof of fertilization in uncleaved human oocytes.
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Cell Division↗
Biomedical subjects
L Mettler
Publications and source records attributed to L Mettler.
Use of a modified test system to determine early pregnancy factor (EPF) levels in patients with normal first trimester pregnancy and after therapeutic abortion.
Human pregnancy can be recognized as early as 48 hours after conception using the rosette inhibition test (RIT) to detect early pregnancy factor (EPF). A modification of the test as originally described by Morton et al. is demonstrated using a monoclonal pan T-cell antibody. Studies were performed to investigate the relationship between EPF levels during normal first trimester intact pregnancies and therapeutic abortions. The change in EPF levels is compared with those of hormones after interrupted pregnancy.
Abortion, Therapeutic↗
[Diagnosis and therapy of immunologic sterility].
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Animals↗
[Effect of various factors on in vitro fertilization (IVF) and embryo transfer in the human in clomiphene/hMG/hCG stimulated cycles].
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.
Chorionic Gonadotropin↗
In vitro fertilization and embryo replacement at the Department of Obstetrics and Gynecology, University of Kiel, F.R.G.
From July 1982 until September 1983, 194 pelviscopies were performed in Kiel (phase I). The patients were stimulated either with Clomid/human menopausal gonadotropin hMG)/human chorionic gonadotropin (hCG) or with Clomid/hCG or hMG/hCG alone. Follicular maturation was monitored by ultrasonography and the daily measured E2 and luteinizing hormone (LH) response. Surgic therapeutical pelviscopy with follicular puncture followed 36 hr after hCG application. Oocytes were incubated in either Ham's F-10 or Menezo B2 medium in an automatically gas-controlled exsiccator. Forty-eight hours after insemination normal-looking four- to eight-cell embryos were replaced into the uterine cavity. Oocytes were successfully collected in 87.4% of the patients, with an average of 2.2 oocytes per patient. Eighteen pregnancies resulted of 101 embryo replacements. The overall pregnancy rate was 16.2% per replacement and 9.3% per pelviscopy. Undivided oocytes and polyploid embryos were analyzed cytogenetically. From October 1983 to October 1984 the overall pregnancy rate after 144 pelviscopies and 88 embryo replacements (phase II) improved to 23.9% per replacement and 14.6% per pelviscopy.
Blastocyst↗
[Pregnancy and birth of monozygous female twins following in vitro fertilization and embryo transfer].
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.
Adult↗
Immunological aspects of pathological pregnancy (infertility immunology).
As, theoretically, any of the many mechanisms that appear to operate in normal pregnancy to maintain maternofetal immunologic balance could be altered in pathologic pregnancy, a detailed study of the known immunological aspects of pathological pregnancy is given. After looking into the parental genetic heterozygosity and fertility, a good explanation of the antifetal immunoresponse by the pregnant mother including the antibody formation is shown. The placental transport versus trapping of immunoglobulins is described in detail and it is also shown that the placenta is a source of a number of agents that inhibit cell-mediated immunity (HCG, HPL and steroids). These substances may affect a local inhibition of maternal cell-mediated immunity in the vicinity of the trophoblast. The placenta serves as an effective barrier for transfer of immunocompetent maternal lymphocytes to the fetus. The paramount role of the trophoblast as a barrier, while itself escaping immunologic destruction, calls for further studies.
Antibody Formation↗
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)
Ejaculation↗
Enzyme-linked immunosorbent assays for sperm antibody detection and antigenic analysis.
Enzyme-linked immunosorbent assays for estimation of antibodies against human sperm and for determination of antigenic reactivity of spermatozoal proteins were established. Sperm immobilized on PHA-coated microtiter plates or solubilized spermatozoal antigens adsorbed on poly(L)-lysine coated microtiter plates were used as the solid phase. Assay of sperm antibodies was performed by incubation of the test samples with the solid phase followed by incubation with anti-Ig conjugated to peroxidase. Sigmoidal antibody dilution curves were obtained with rabbit and mouse anti-sperm sera. The ELISA was effectively used to screen production of anti-sperm antibodies by mouse myeloma x splenocyte hybridomas. The sensitivity of this ELISA for sperm antibodies was more than 1000-fold greater than the classical tray sperm immobilization test, and was comparable in sensitivity to a radioimmunoassay using 125I-labeled protein A as the tracer. Sperm immobilized on PHA-coated plates exhibited significantly greater antigenic reactivity in both the ELISA and RIA compared with methanol fixed sperm. In a competitive inhibition ELISA, linear Logit-log dose-response curves were obtained with detergent solubilized spermatozoal antigens. The assay was used to monitor the purification of the solubilized spermatozoal antigens by chromatofocussing; a more than 60-fold increase of antigenic potency of purified sperm antigen compared with unfractionated sperm extract was evident in the competitive ELISA.
Animals↗
In vitro studies of ovarian function in the isolated perfused rabbit ovary.
An in vitro model for studies of preovulatory follicle and ovulation is described, using a recirculating system for perfusion of isolated rabbit ovaries. The preovulatory follicle was studied by addition of human chorionic gonadotropin (hCG) to the perfusion medium 1.5 h after the start of perfusion. Ovulations occurred approximately 13 h after in vivo hCG injection to the animal and 5-10 h after the start of perfusion. The ultrastructural changes during the perfusion were similar to in vivo conditions. The release of estradiol and progesterone into the medium are consistent with in vivo measurements. This in vitro model is probably useful for further studies of the biochemistry and morphology of ovarian function.
Animals↗
Three step medical and surgical treatment of endometriosis.
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Danazol↗
[Hybridoma technic: a new biological dimension in reproduction research].
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Animals↗
Identification of a sperm receptor-glycoprotein from the porcine zona pellucida.
Porcine zona pellucida were solubilized by treatment with lithium 3,5-diiodosalicylate. Following further purification by phenol/water extraction, ethanol precipitation and high performance liquid chromatography a total of 11 fractions were obtained. Fractions in the molecular mass range, that were previously found to possess antigenic properties, were tested in a spermatozoa radio-receptor binding assay. One of the fractions with an N-terminal amino-acid residue of alanine was found to have spermatozoa receptor properties. This result might aid future studies on the spermatozoazona pellucida interactions at the molecular level.
Amino Acids↗
Estimation of anti-spermatozoa antibody concentrations by a [125I]protein-A binding assay in sera of infertile patients.
A solid-phase radioimmunoassay was developed for measuring the concentration of antispermatozoa antibodies in the sera of sterile patients. Whole human spermatozoa fixed in methanol and immobilized on plastic microtiter plates were used as antigens, and the spermatozoa-bound antibodies were quantitated with the aid of [125I]protein-A. The assay is specific, simple to perform and up to 1000 tests can be carried out conveniently by one technician on a single working day. The new test was shown to correlate better with immunological sterility than the sperm microagglutination and immobilisation tests. An antispermatozoa antibody concentration of 210 micrograms/l and above was found be indicative of an immunological cause of infertility.
Agglutination Tests↗
[The importance of sperm quality for in vitro fertilization--results of studies in human systems and zona pellucida-free hamster cell system].
In this report the minimum requirements for ejaculates to be used for in vitro fertilization of human oocytes are defined. Based on the 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 spermio-cytogram. The total volume of the ejaculate should not be less than 1 ml. Samples for insemination of human oocytes must be proven to be free of pathological microorganisms. In more than 40% of the cases examined bacteria concentrations of more than 10(5)/ml were detected during routine bacteriological check-ups. Especially enterococci und mycoplasma were frequently found. To further establish the quality of human spermatozoa, 91 ejaculate probes were tested within the hamster system using zona-free oocytes. A statistically significant correlation between sperm quality and the penetration rates of human spermatozoa into zona-free hamster oocytes was shown. A positive effect on penetration rates was observed after addition of L-ascorbic acid and Douglas-aspirate to the incubation medium. Rates increased 12 and 17%, respectively. Addition of kallikrein showed no effect on the penetration rates. Application of ejaculates with high concentrations of leucocytes for insemination of hamster oocytes resulted in clearly reduced penetration rates. Leucocyte concentrations of more than 10% led to penetration rates of 0%. The same reduction was observed when ejaculates with high concentrations of pathological micro-organisms (more than 10(5)/ml) were tested. After addition of sperm antibody containing sera from female sterility patients to the incubation medium also clear reduction of penetration rates was evident depending on: the antibody concentration, the initial motility of the spermatozoa used, and on the level of the primary penetration rate obtained without addition of antiserum.
Animals↗
Immunogenic potency of the zona pellucida.
New Zealand white rabbits were immunized with low doses of pig zona pellucida material with the aim of reducing nonspecific antibodies in the antiserum. The antibody levels were assayed by the standard precipitation and immunofluorescence methods. The titers produced were comparable with those obtained using large amounts of zona material.
Animals↗
Rosette inhibition test: a multicentre investigation of early pregnancy factor in humans.
The rosette inhibition test for the detection of early pregnancy factor is described in detail. The extended methodology presented here represents the cumulative experience of three independent laboratories. Special reference is made to the effect on the assay of varying the conditions of rosette formation between lymphocytes and sheep red blood cells. Antilymphocyte sera prepared for use in the rosette inhibition test fell into three categories: (i) with no rosette inhibiting activity, (ii) with rosette inhibiting activity which is not affected by the presence of EPF, and (iii) rosette inhibiting activity which is significantly increased in the presence of EPF. To date, this third reaction has been found to be a specific indication of the presence in serum of early pregnancy factor.
Animals↗
The frequency of serological anti-zona pellucida activity in males, females and children.
945 human sera from children, fertile and infertile females, pregnant women and fertile and infertile males were tested for antibodies to porcine zona pellucida by immunofluorescence and the pronase-induced zona-lysis inhibition test. Positive sera were found in each of the groups examined, with a higher percentage of anti-zona activity in infertile, pregnant and menopausal women. There was no correlation between anti-sperm antibody titer and the incidence of anti-zona activity in either female or male human sera, nor did the occurrence of anti-zona activity correlate with a particular blood group type.
Adolescent↗