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Biomedical subjects

L Veeck

Publications and source records attributed to L Veeck.

At least 19 recordsLinked to original sources
PubMed1999

Autologous endometrial co-culture in patients with repeated failures of implantation after in vitro fertilization-embryo transfer.

PURPOSE: Our purpose was to evaluate the effect of coculture on preembryo development and clinical outcome. METHODS: Enrolled patients underwent a luteal-phase endometrial biopsy. The tissue was then enzymatically digested (collagenase) and the stromal and glandular cells were separated by differential sedimentation rates. These cells were cultured to confluence, released, and then cryopreserved until the patient's in vitro fertilization (IVF)-embryo transfer (ET) cycle. All normally fertilized oocytes were then placed on the co-cultured cells until transfer on day 3. Preembryo development on co-culture was compared to that in the patient's noncocultured previous cycle. Implantation and clinical pregnancy rates were compared to those in a control group of patients undergoing IVF during the study period who were matched for age, stimulation protocol, number of oocytes retrieved, and preembryos transferred. RESULTS: Twenty-nine women underwent 31 cycles of IVF-ET. On day 3 the overall mean number of blastomeres per preembryo on co-culture compared to that in the patient's previous cycle was 6.3 +/- 1.8 vs. 5.6 +/- 1.2 (P = 0.04). The average percentage of cytoplasmic fragments on co-culture compared to the previous cycle was 16 +/- 9% vs. 19 +/- 9% (P = 0.32). At transfer, after preembryo selection, the mean number of blastomeres per preembryo on co-culture compared to that in the patient's previous cycle was 6.8 +/- 1.6 vs. 6.6 +/- 1.3 (P = 0.5). The implantation and clinical pregnancy rates between co-culture and the matched control group were 15% (14/93) vs. 13% (16/124) (P = 0.79) and 29% (9/31) vs. 25% (10/40) (P = 0.45). CONCLUSIONS: There was a significant improvement in the average number of blastomeres per preembryo on co-culture compared to that in the patient's previous noncoculture cycle. The overall implantation and clinical pregnancy rates between co-culture and a matched control group were not significantly different.

Adult↗
PubMed1998

Human preembryo development on autologous endometrial coculture versus conventional medium.

OBJECTIVE: To evaluate the effect of autologous endometrial coculture versus conventional medium on preembryo development. DESIGN: Controlled systematic clinical study. SETTING: University-based IVF center. PATIENT(S): Women with a history of failed IVF-ET with poor preembryo quality. INTERVENTION(S): Patients underwent a luteal phase endometrial biopsy. The tissue then was digested enzymatically, and the stromal and glandular cells were separated by differential sedimentation rates. These cells were cultured to confluence, released, and then cryopreserved until the patient's IVF-ET cycle. All normally fertilized oocytes then were allocated systematically to growth on autologous endometrial coculture or conventional medium until transfer on day 3. MAIN OUTCOME MEASURE(S): Preembryo blastomere numbers and cytoplasmic fragmentation rates were measured. RESULT(S): Forty-two women underwent 44 cycles of IVF-ET. In the morning on day 3, the mean (+/-SD) number of blastomeres and cytoplasmic fragments per preembryo on coculture compared with conventional medium was 5.9+/-1.5 versus 5.5+/-1.4 and 21%+/-13% versus 24%+/-11. At transfer the mean (+/-SD) number of blastomeres per preembryo on coculture was 7.4+/-1.3 versus 6.7+/-1.9 on conventional medium. CONCLUSION(S): There was a significant improvement in the mean (+/-SD) number of blastomeres per preembryo and decrease in the fragmentation rate for preembryos on autologous endometrial coculture compared with noncocultured preembryos from the same patient.

Adult↗
PubMed1998

Comparison of clinical outcome after cryopreservation of embryos obtained from intracytoplasmic sperm injection and in-vitro fertilization.

The impact of intracytoplasmic sperm injection (ICSI) on cryopreserved zygotes and embryos was evaluated by comparing embryo survival and implantation between embryos derived from ICSI and those derived from standard insemination procedures. The study included patients whose excess zygotes and embryos were cryopreserved between September 1993 and December 1994 and who subsequently underwent a frozen embryo transfer. Embryo survival, clinical pregnancy rates per transfer and pregnancy outcome were compared. Three hundred and thirty eight cryopreservation cycles, during which 1471 embryos were cryopreserved, were included in this study. Of those, 961 were derived from oocytes fertilized by insemination in vitro and 510 were derived from oocytes fertilized by ICSI. A total of 690 of the embryos (451 in the insemination group and 239 in the ICSI group) have since undergone a thaw cycle. The embryo survival rates were similar between the two groups (70.5 and 73.2%, insemination and ICSI respectively) and were not significantly affected by the stage at cryopreservation. There was no significant difference in pregnancy rates per transfer (31.8 and 32.3%), the preclinical pregnancy loss rate (16.7 and 23.8%), or the clinical miscarriage rate (16.7 and 23.8%) between the insemination and the ICSI groups respectively. It is concluded that ICSI does not have an adverse impact on the survival and successful implantation of cryopreserved and thawed embryos.

Cryopreservation↗
PubMed1998

Granulocyte macrophage-colony stimulating factor production by autologous endometrial co-culture is associated with outcome for in vitro fertilization patients with a history of multiple implantation failures.

PROBLEM: To determine whether granulocyte macrophage (GM)-colony stimulating factor (CSF) produced by autologous endometrial co-culture was associated with outcome in 53 patients with a history of multiple in vitro fertilization failures. METHOD OF STUDY: The conditioned media from endometrial co-culture cells exposed or non-exposed to human embryos was analyzed for GM-CSF. RESULTS: Exposure or non-exposure to an embryo did not result in an enhancement of GM-CSF production. Insignificant levels of GM-CSF were determined from media alone. ROC analysis revealed that levels of GM-CSF from supernatants of endometrial co-culture exposed to embryos that measured below 130 pg/ml reflected a diminished prognosis (5/17 had a positive pregnancy vs. 21/36 with GM-CSF levels greater than 130 pg/ml; P < 0.05). CONCLUSIONS: The improved outcome associated with GM-CSF values greater than 130 pg/ml may reflect: 1) a direct positive effect of GM-CSF; 2) an embryotrophic factor upregulated by GM-CSF; or, 3) that GM-CSF functions as a marker for the importance of the glandular component in endometrial co-culture systems.

Adult↗
PubMed1995

Intracytoplasmic sperm injection: achievement of high pregnancy rates in couples with severe male factor infertility is dependent primarily upon female and not male factors.

OBJECTIVE: To determine the efficacy and factors affecting outcome of intracytoplasmic sperm injection (ICSI) in patients with severe male factor infertility. DESIGN: Prospectively designed clinical trial of patients selected to participate in the study based upon the following inclusion criteria: previous total failed fertilization or unsuitable sperm parameters for conventional IVF. SETTING: Tertiary care academic center. PATIENTS: Ninety-two consecutive couples undergoing IVF therapy augmented with ICSI during April through December 1994 were studied. MAIN OUTCOME MEASURES: Fertilization and ongoing implantation and pregnancy rates (PRs). RESULTS: A total of 1,163 preovulatory oocytes were manipulated, yielding a diploid fertilization rate of 60.9%; the oocyte damage rate was 13.2%. The transfer rate was 95% with 43.1% of cycles having excess embryos that were cryopreserved. Overall, the clinical and ongoing PRs per transfer were 31.9% and 26.8%, respectively. None of the sperm parameters of the original semen analysis correlated with ICSI outcome. Female age did not affect fertilization results but had a significant impact on PR (< 34 years: 48.9%; 35 to 39 years: 22.9%; > or = 40 years: 5.9% clinical PR per transfer). CONCLUSIONS: Intracytoplasmic sperm injection offers a new and powerful therapeutic option to treat couples with severe male factor infertility associated with a variety of sperm abnormalities. An adequate female age is a pivotal factor determining a successful outcome.

Adult↗
PubMed1992

Preclinical models for human pre-embryo biopsy and genetic diagnosis. II. Polymerase chain reaction amplification of deoxyribonucleic acid from single lymphoblasts and blastomeres with mutation detection.

OBJECTIVE: To demonstrate the use of the polymerase chain reaction in the amplification of deoxyribonucleic acid (DNA) from single human lymphoblasts and mouse blastomeres. Amplified target genes for diagnosis of sickle cell anemia and Tay-Sachs are shown. Similarly, the sparce fur mouse model for ornithine transcarbamylase deficiency was used as an X-linked system for demonstration of mutation detection after biopsy of a single blastomere. A new diagnostic method for the detection of the ornithine transcarbamylase mutation using the restriction enzyme Mse I is presented. Accuracy and reproducibility were assured. DESIGN: Polymerase chain reaction proficiency test for amplification from single cells was studied. Also, accuracy of mutation detection systems was demonstrated. SETTING: Laboratories of The Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School. PATIENTS, PARTICIPANTS: We used the sparse fur mouse model and human blood cells. INTERVENTIONS: Pre-embryo biopsy, polymerase chain reaction amplification, and mutation detection were performed. MAIN OUTCOME MEASURES: Accuracy and reproducibility of DNA amplification without contamination, as well as efficient diagnostic analysis, from both single somatic and embryonic cells were shown. RESULTS: DNA amplification from single cells was uniformly rapid (6 to 10 hours) reproducible (n = 220) and accurate (n = 52). CONCLUSIONS: Our findings support the feasibility of clinical application for pre-embryo biopsy and genetic diagnosis of specific heritable diseases.

Amino Acid Sequence↗
PubMed1992

Successful fertilization and pregnancy outcome in in-vitro fertilization using cryopreserved/thawed spermatozoa from patients with malignant diseases.

Cryopreservation of spermatozoa before treatment is the only proven effective method available to circumvent the sterilizing effect of therapy in some patients with malignant diseases. Because of impaired sperm quality after freezing and thawing in-vitro fertilization/embryo transfer (IVF/ET) was indicated in 10 patients (12 cycles) during 1986-1990. The patient's mean age was 33.4 +/- 1.6 years. The following diagnoses were made: seminoma (1), testicular carcinoma (3), leiomyosarcoma of the prostate (1), Wegener's granulomatosis (1), non-Hodgkin's (1) and Hodgkin's lymphoma (3). When motile spermatozoa could be recovered after thawing, the total fraction of motile spermatozoa after swim-up separation ranged from 0.2 to 4.2 x 10(6) spermatozoa/ml (eight patients, nine cycles). In all these cases, insemination was performed with multiple oocytes per dish. Fertilization was achieved when swim-up recovered a mean of 1.8 +/- 0.5 x 10(6) spermatozoa/ml and when insemination was performed with at least a calculated concentration of motile spermatozoa of 1 x 10(5) spermatozoa/oocyte. The fertilization rate of preovulatory oocytes was 60%. Four patients achieved a pregnancy: two of them delivered a single healthy baby, one delivered triplet healthy babies and one had a preclinical abortion. In two patients (three cycles), no motile spermatozoa were recovered after thawing, and micromanipulation of oocytes for assisted fertilization was performed. Although fertilized oocytes were transferred, those couples did not achieve a pregnancy. Patients with lymphopathies had the best results, whilst those with testicular neoplasms had the poorest outcome, thus suggesting a poor gametogenic function in the non-affected testis. These results give hope to some patients with malignant diseases to maintain their reproductive capacity through sperm banking and IVF/ET.

Adult↗
PubMed1991

Human preovulatory oocytes have a higher sperm-binding ability than immature oocytes under hemizona assay conditions: evidence supporting the concept of "zona maturation".

OBJECTIVE: The purpose of this study was to investigate the sperm-binding potential of human oocytes at different stages of nuclear maturation under hemizona assay (HZA) conditions. DESIGN: This was a prospective study designed in a blinded fashion. SETTING: Academic research environment approved by the Institutional Review Board. PATIENTS: Surplus oocytes, donated by patients undergoing in vitro fertilization therapy after gonadotropin stimulation, were analyzed. Semen from a fertile donor was used in all assays. INTERVENTIONS: Five groups of oocytes were considered: (1) immature, prophase I; (2) metaphase I; (3) metaphase II; (4) inseminated, unfertilized metaphase II; and (5) immature, prophase I oocytes matured in vitro to metaphase II. Oocytes were stored in salt solution (pH 7.2) and microbisected before assay. MAIN OUTCOME MEASURE(S): Tight binding of sperm to the zona pellucida under HZA conditions was evaluated after 4 hours of gametes coincubation. RESULTS: Metaphase II oocytes (groups 3 and 4) had significantly higher binding than other groups (P = 0.0001). The mean value of the difference between the two halves (hemizona) was not significant, thus showing a small intra-assay variation for all maturational stages. CONCLUSIONS: Full meiotic competence of human oocytes is associated with an increased zona pellucida-binding potential.

Double-Blind Method↗
PubMed1991

Analysis of ectopic pregnancies resulting from in-vitro fertilization and embryo transfer.

The purpose of this study was to analyse the risk factors, stimulation characteristics, site and outcome of pregnancy and future fecundity of patients who develop ectopic pregnancies after in-vitro fertilization (IVF). Of 3145 transfer cycles between January 1981 and July 1989, 27 (3.3%) of the resulting 825 pregnancies were ectopic. There was a significantly greater incidence of a prior ectopic pregnancy in the study group compared to the controls. Compared to matched controls with intrauterine pregnancies, the study group had significantly higher peak oestradiol levels. Twenty-one ectopic pregnancies were ampullary, two were interstitial, one was abdominal, one was cervical and two were heterotopic. Sixteen of the patients subsequently underwent 40 IVF attempts with a pregnancy rate of 28% per transfer. We conclude that patients with a prior ectopic pregnancy are at risk for an IVF ectopic pregnancy. The subsequent IVF outcome of those who develop ectopic pregnancies after IVF is encouraging.

Adult↗
PubMed1988

Mouse embryo quality control for toxicity determination in the Norfolk in vitro fertilization program.

Ongoing quality control is necessary as part of the maintenance and improvement of a successful human in vitro fertilization (IVF) program. Using a mouse quality-control culture system, several instrument preparation protocols were reevaluated to determine their efficiency in the control or elimination of potential toxicity. Dilute concentrations of urine and endometrial fluid were also tested. Medium rinsed through laparoscope and aspiration needles failed to support embryo development. This effect was reversed in needles that were pretreated with rinses of Dulbecco's phosphate-buffered saline. Endometrial fluid demonstrated no obvious toxic effect, but urine-exposed embryos arrested in the two-cell state. The importance of periodic evaluation of materials and their pretreatment before use in in vitro fertilization of human oocytes is essential to ensure control of potentially toxic substances.

Animals↗
PubMed1988

Corrective measures and pregnancy outcome in in vitro fertilization in patients with severe sperm morphology abnormalities.

Sperm morphology evaluated by new, strict criteria is a good predictor of outcome in in vitro fertilization (IVF). This study aimed (1) to determine whether the fertilization rate of preovulatory oocytes in patients with abnormal morphology can be improved by increasing insemination concentration at the time of IVF and (2) to evaluate the pregnancy outcome in patients with abnormal sperm morphology. Three groups were studied: (1) normal morphology, (2) good prognosis pattern, and (3) poor prognosis pattern. All other sperm parameters were normal. Group 3 had a lower overall fertilization rate, lower pregnancy rate/cycle, and lower ongoing pregnancy rate/cycle. Groups 2 and 3 showed a higher miscarriage rate, although not significantly different from group 1. By increasing insemination concentration from 2- to 10-fold, the fertilization rate in group 3 increased from 14.5% to 62.6%. However, pregnancy outcome did not improve. We conclude that patients with severe sperm head abnormalities have a lower ability to establish successful pregnancies, even though fertilization may be achieved.

Abortion, Spontaneous↗
PubMed1987

Kinetic immunonephelometric determination of protein concentrations in follicular fluid.

Nonsteroidal biochemical markers of oocyte maturity could presumably aid in oocyte selection and in the timing of insemination for in vitro fertilization. We assessed the usefulness of six potential markers found in follicular fluid. We used kinetic immunonephelometry to measure concentrations of alpha 1-antitrypsin, alpha 2-macroglobulin, antithrombin III, ceruloplasmin, fibrinogen, and plasminogen in 53 fluids and the corresponding plasma from 20 women undergoing in vitro fertilization. Specimens were obtained from both mature and immature follicles from each woman. The respective protein concentrations in follicular fluid could be assayed with a between-assay CV of 1.5% to 3.2%. Analytical-recovery studies indicated that only fibrinogen and antithrombin III concentrations were altered (by 120% and 75%, respectively) during aspiration of the follicle into saline. All protein concentrations were significantly increased in mature follicles, as were the follicular fluid:plasma concentration ratios for alpha 1-antitrypsin, antithrombin III, and ceruloplasmin.

Antithrombin III↗
PubMed1987

Follicular fluid estradiol and progesterone are markers of preovulatory oocyte quality.

Follicular fluid estradiol (FF E2) and progesterone (P) have previously been shown to vary with the maturation of the follicle. A profile of follicular fluid E2 and P was obtained from follicles of 19 preovulatory oocytes, which resulted in normal term pregnancies from single embryo transfers. A narrow band was defined around the linear regression line E2/P = 1.2 X 10(-2) + 1.57 X 10(-4) E2, r = 0.806, P less than 0.01, when E2/P was plotted versus E2 that included 84.2% of these oocytes. Eleven of the 19 pregnancies were clustered between an E2 of 240 to 460 ng/ml and an E2/P ratio of 0.044 to 0.084. Sixty-eight patients undergoing in vitro fertilization and embryo transfer were studied. A significantly greater number of preovulatory follicles from the fertilized, cleaved, and pregnant group (10 of 40) was found in the cluster than from the fertilized, cleaved, but not pregnant group (4 of 46; P = 0.04) and fertilized but not cleaved group (0 of 13; P = 0.04). Significantly more pregnant patients (14 of 20) had a preovulatory follicle within the band than nonpregnant patients (11 of 36) (P less than .02), as well as in the cluster (10 of 20), compared with 4 of 36, respectively (P less than 0.01). It may be concluded that a range of FF E2 and E2/P values define a stage of maturation that is compatible with successful pregnancies.

Estradiol↗
PubMed1985

Male factor evaluation in in vitro fertilization: Norfolk experience.

Thirty-three patients from the in vitro fertilization (IVF) program at Norfolk are critically reviewed. A battery of tests was designed and an endocrine investigation was carried out on these patients. The fertilization rate for preovulatory oocytes was lower than in the normal male population (39.6% versus 88.6%). When total concentration of sperm with rapidly progressive motility was less than 6 X 10(5), to fertilize several eggs together the fertilization rate was zero. No fertilization was obtained when the number of sperm with rapidly progressive motility recovered after the separation was less than 1.5 X 10(6). The hamster zona-free oocyte penetration test correlated well with the human IVF system. The other parameters investigated did not show good correlation. When fertilization was achieved, the results of the IVF procedure in the series reviewed rendered a 30.8% pregnancy rate per transfer in 26 transfers. Fifty percent of the pregnancies were normal (either ongoing or delivered). Thirty-seven percent were preclinical miscarriages, and 12.5% were clinical abortions. In the abnormal male population, higher concentrations of sperm per egg should be used for insemination for achievement of optimum fertilization rates. Once fertilization is obtained, the results do not differ substantially from the IVF population at large.

Animals↗
PubMed1984

In vitro fertilization in Norfolk, Virginia, 1980-1983.

Three years of progress of the Vital Initiation of Pregnancy (VIP) Program in Norfolk is reported. No conception resulted from 41 oocyte aspirations during spontaneous menstrual cycles in 1980. An average of 3.7 oocytes per cycle, or a 73.5% recovery rate, resulted in 362 human menopausal gonadotropin/human chorionic gonadotropin-induced cycles from January 1981 to March 1983. Forty percent of the oocytes recovered from these cycles were preovulatory, 35% atretic, and 25% immature. Immature oocytes were often matured in vitro, fertilized, and found to produce pregnancies. A total of 62 pregnancies occurred, which represents a 17 or 23% pregnancy rate, based on laparoscopies or embryo transfers, respectively. There were 11 preclinical and 7 clinical miscarriages. Twenty-nine normal babies have been delivered, including a set of twins. The remainder appears to be normally progressing pregnancies. Polyspermia was observed in 8.8% of the fertilizable oocytes.

Adult↗
PubMed1984

Correlation of human menopausal gonadotropin/human chorionic gonadotropin stimulation and oocyte quality in an in vitro fertilization program.

One hundred forty-seven cycles in normal ovulatory patients are reported. All were stimulated with human menopausal/human chorionic gonadotropin. Three estrogen responses were identified: normal, high, and low. Patients who achieved pregnancy formed a fourth category, the pregnancy group. The number of preovulatory and immature oocytes, the preovulatory and immature oocytes that fertilized normally or abnormally, the ones that cleaved in culture, and the ones that were transferred were used as parameters to compare quality of the oocytes in each of the estrogen responders. No significant differences were found in any of them. Abnormal zonae pellucidae are described as possibly due to overmaturation of the follicle. No significant difference in the proportion of abnormal zonae in the different categories was found.

Adult↗