Cryopreservation of embryos and pregnancy rates after IVF.
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Adult↗
Biomedical subjects
B A Lieberman
Publications and source records attributed to B A Lieberman.
Endometrium in in-vitro fertilization cycles: morphological and functional differentiation in the implantation phase.
Secretory differentiation of endometrium after multiple follicular stimulation using gonadotrophin releasing hormone and human menopausal gonadotrophin has been studied both histologically and immunohistochemically in 30 women undergoing in-vitro fertilization treatment. None had embryo transfer. Patients were randomly allocated to receive luteal phase support with a single dose of human chorionic gonadotrophin. The latter failed to produce any significant enhancement of endometrial structure or secretions. Appropriate glandular morphology was present in a greater proportion of those who were successfully stimulated than those who responded poorly. However, defective secretion of the cycle-dependent component studied, using monoclonal antibody D9B1, was demonstrated in two-thirds of cases regardless of the ovarian response. Early vascular maturation in the stroma was a common finding, and was thus considered as a feature of structural modulation of these endometria.
Antibodies, Monoclonal↗
The use of cryopreserved aged human oocytes in a test of the fertilizing capacity of human spermatozoa.
The use of cryopreserved aged human oocytes in a diagnostic test of sperm fertilizing ability was evaluated. Oocytes arising from assisted conception cycles and showing no signs of fertilization 48 h post-insemination were cryopreserved by one of two methods. An ultrarapid method using dimethyl sulphoxide gave poor post-thaw results, with only 5/69 (7.2%) oocytes surviving. Oocytes frozen by a slow method using propanediol as the cryoprotectant gave better survival rates (359/594; 60%). Fertilization by donor spermatozoa of these thawed oocytes was poor (15/63; 24%) when the zona pellucida was left intact. To improve this, the zona was enzymatically removed using pronase. These zona-free oocytes were then inseminated with spermatozoa from a fertile donor or from men previously exhibiting fertilization failure in an in-vitro fertilization treatment cycle. The fertilization rate in the patient group (41/91; 45%) was significantly lower than in the donor group (16/18; 89%) (P less than 0.02). There was also a significant (P less than 0.03) reduction in the median number of pronuclei per oocyte (2.9 versus 4.5). These results show that aged oocytes can be effectively cryopreserved to establish a bank for use in a test to identify men with impaired sperm fertilizing capacity.
Cell Survival↗
Clinical and endocrinological changes in women following ovulation induction using buserelin acetate/human menopausal gonadotrophin augmented with biosynthetic human growth hormone.
Biosynthetic human growth hormone added to an ovarian stimulation regime of human menopausal gonadotrophin (HMG) for IVF treatment improves the response of women who were previously resistant. This study investigated the efficacy of growth hormone (GH)/buserelin/HMG treatment in women with a previous normal response to buserelin/HMG stimulation. Ten patients (28-36 years, mean 32.5 years) were treated with GH (6 IU/day) plus buserelin/HMG. A control group of 10 women (28-37 years mean 31.0 years) received buserelin/HMG alone. All were given buserelin 500 micrograms and 2 ampoules (150 IU) HMG daily once pituitary suppression had been confirmed. There was no improvement in the GH group as assessed by follicular growth rate or number, oocyte number per woman and pregnancy rate. There was no effect of GH upon the serum oestradiol level and the follicular fluid levels of oestradiol, GH and inhibin. Serum IGF-1 increased significantly during GH administration, returning to pre-treatment levels 2 days after the last dose of GH. Follicular IGF-1 was much higher in the GH-treated group than the controls. Significant correlations were found in the GH-treated group between follicular fluid GH and follicular fluid oestradiol concentrations and between follicular GH and follicular size. Follicular IGF-1 was correlated with the serum IGF-1 concentration on day 8 of the GH/HMG treatment. In conclusion GH/buserelin/HMG treatment in women with a previous normal response to buserelin/HMG stimulation increased their serum and follicular IGF-1 concentrations. However, it does not improve the clinical ovarian response or the follicular secretion of oestradiol or inhibin.
Adult↗
Outcome of treatment subsequent to the elective cryopreservation of all embryos from women at risk of the ovarian hyperstimulation syndrome.
From 1st June 1989 to 31st May 1991, 78 women with a serum oestradiol level greater than 3500 pg/ml on the day of the ovulatory trigger, following pituitary suppression with buserelin and ovarian stimulation with human menopausal gonadotrophins (HMG), had all their embryos electively cryopreserved at the pronucleate stage to minimize the risk of developing ovarian hyperstimulation syndrome (OHS). Treatment with buserelin was continued in the luteal phase. A median of 19 oocytes (range 7-43) was obtained and 12 embryos (range 1-37) frozen per cycle. Twenty-one (27%) women developed OHS (six severe). Women developing OHS had higher (P less than 0.05) serum oestradiol concentrations on the 7th day after oocyte retrieval, compared to those who did not. No differences were found for any of the following criteria: aetiology of infertility, age, total dose of HMG, number of oocytes, fertilization rate or freeze-thaw survival of embryos. Subsequently, 125 frozen-thawed embryo replacements have been undertaken, using buserelin and hormone replacement therapy (HRT) (n = 93) or natural cycles (n = 32). The overall freeze-thaw survival and implantation rates per embryo were 71.8 and 11.7%, respectively. The pregnancy rates in natural cycles (19%) and buserelin/HRT cycles (29%) were not significantly different.
Buserelin↗
Is continuation of a gonadotrophin-releasing hormone agonist (GnRHa) necessary for women at risk of developing the ovarian hyperstimulation syndrome?
A total of 28 women scheduled for in-vitro fertilization used buserelin and human menopausal gonadotrophin (HMG) for ovarian stimulation. One group (I) of 17 women was given human chorionic gonadotrophin (HCG 10,000 IU) to trigger ovulation, but the resulting embryos were electively cryopreserved because of the risk (serum oestradiol greater than or equal to 3500 pg/ml) of developing the ovarian hyperstimulation syndrome (OHSS). Six women continued the buserelin therapy in the luteal phase and eleven did not. In group II (n = 11), the HMG injections were discontinued because of an exaggerated ovarian response and the HCG was omitted. Six of these women continued the buserelin injections until the onset of menses and five did not. In both groups, the ovarian response to induction of ovulation (serum oestradiol concentrations and number of follicles) was similar for those who did or did not continue buserelin therapy. There was no difference in the rate of ovarian quiescence (weekly fall in serum oestradiol concentration following the stimulation) between those women who did or did not continue the buserelin therapy in either group. The serum luteinizing hormone concentrations remained low in all women in both groups. We conclude that the omission of buserelin therapy after discontinuing the HMG in women at risk of developing OHSS does not affect subsequent ovarian quiescence.
Adult↗
Pregnancy following intravaginal embryo transportation.
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Adult↗
Role of the ovary in the synthesis of placental protein-14.
Women with absent ovarian function provide an opportunity to investigate the ovarian contribution to secretion of placental proteins (PP), such as PP14, in early pregnancy. We present data on serum PP14 levels in 12 women (median age, 30.5 yr; range, 26-37 yr) with premature ovarian failure (POF) who conceived after ovum donation and embryo transfer with exogenous sex steroid support using transdermal (n = 5) or oral (n = 7) estradiol and vaginal (n = 8) or im (n = 4) progesterone. The women were closely monitored throughout early pregnancy, with measurement of serum levels of estradiol (E2), progesterone (P4), and PP14. Levels of E2 and P4 were entirely normal. Levels of PP14 were significantly subnormal (P = 0.008) in all 12 agonadal women compared with levels of PP14 in a control group of women with normal ovarian function between 6-12 weeks gestation. Basal and peak levels for subjects with absent ovarian function were 40 and 124 micrograms/L, respectively. For each week between 6-12 weeks of pregnancy, the mean serum levels of PP14 for women with normal ovarian function were between 706-940 micrograms/L. These observations support the concept that PP14 arises from the ovary in early pregnancy or that factors under the control of the maternal ovary are involved in its production by the endometrium.
Adult↗
Cryopreservation of human embryos at the pronucleate, early cleavage, or expanded blastocyst stages.
Supernumerary embryos following treatment by IVF or GIFT were cryopreserved at the pronucleate, early cleavage or expanded blastocyst stages. The success of embryo cryopreservation at these stages was evaluated in terms of (i) the proportion of embryos surviving the freeze/thaw procedure; (ii) the proportion of patients reaching embryo replacement; and (iii) the incidence of pregnancy per replacement. Significantly more embryos survived when frozen/thawed at the pronucleate (44/61; 72%) or early cleavage stages (48/80; 60%), than at the expanded blastocyst stage (13/34; 38%). A significantly higher proportion of patients had embryo replacements when embryos were frozen/thawed at the pronucleate (17/19; 89%) or early cleavage stages (21/24; 88%), than at the expanded blastocyst stage (9/17; 53%). Following replacement of frozen/thawed pronucleate and early cleavage stage embryos, clinical pregnancy rates of 8/17 (47%) and 3/21 (14%) clinical pregnancies were achieved, respectively. No pregnancies were achieved following replacement of frozen/thawed expanded blastocysts.
Blastocyst↗
The use of biosynthetic human growth hormone to augment ovulation induction with buserelin acetate/human menopausal gonadotropin in women with a poor ovarian response.
The present study has demonstrated the usefulness of GH in augmenting buserelin acetate/hMG to stimulate the rate of growth of follicles in women regarded as poor responders. The PR achieved was extremely encouraging in a group of patients whose prognosis was otherwise poor. Further studies are required to confirm these preliminary data.
Adult↗
High-dose intravenous immunoglobulin in the prenatal management of neonatal alloimmune thrombocytopenia.
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Adult↗
Serum inhibin as an index of ovarian function in women undergoing pituitary suppression and ovarian stimulation in an in vitro fertilization program.
Serum inhibin concentrations, determined by radioimmunoassay, were measured in women undergoing pituitary suppression with buserelin and subsequently ovarian stimulation with human menopausal gonadotropin (hMG). Three types of treatment cycle were investigated, namely (1) cycles showing a normal ovarian response and resulting in a pregnancy; (2) cycles showing a poor response to the hMG, and (3) cycles previously showing a poor ovarian response but augmented with biosynthetic human growth hormone. Good correlations were seen between serum inhibin concentrations and serum progesterone in the luteal phase prior to buserelin (Rs = 0.68), serum oestradiol on the 8th day of hMG administration (Rs = 0.82) or the day of ovulatory trigger (Rs = 0.78), and the number of follicles greater than or equal to 14 mm on the day of ovulatory trigger (Rs = 0.71). These results show inhibin to be a good index of ovarian function in women exhibiting a range of ovarian responses to stimulation in an in vitro fertilization program.
Adult↗
Characterization and functional properties of the A and B forms of human progesterone receptors synthesized in a baculovirus system.
Human progesterone receptors (PR) were overexpressed in Spodoptera frugiperda (Sf9) insect cells using a recombinant baculovirus system. Recombinant viruses were constructed that produced either full-length A (94K) or B (120K) forms of human PR, and each was expressed as a functional protein. Steroid and DNA binding activities were found to be indistinguishable from that of endogenous human PR in T47D breast cancer cells. Moreover, as analyzed by gel-mobility shift, recombinant PR-A and PR-B each bound to specific progesterone response elements in a strictly hormone-dependent manner. Native receptors expressed in Sf9 cells also exhibited structural properties similar to that of endogenous PR. Cytosolic PR (PR-A or PR-B), prepared in low salt buffer, sedimented on density gradients as an 8S oligomeric complex that was converted largely to 4S by treatment with 0.4 M NaCl. Immune isolation of the 8S cytosol PR complex and analysis of protein composition revealed the presence of two specific copurifying proteins of approximately 90K and 70K. The 90-K component was identified immunologically as heat shock protein 90. The 70-K component was not identified but is likely to be the insect equivalent of heat shock protein 70. Immune isolation of PR from Sf9 cells metabolically labeled with [32Pi], revealed that expressed PR was capable of being phosphorylated in insect cells. Hormone addition to Sf9 cells, however, did not stimulate the same increase in PR phosphorylation or upshift in mobility on sodium dodecyl sulfate gels that occurs with endogenous receptors in T47D cells. Thus some, but not all, phosphorylations occur with human PR expressed in Sf9 cells. These phosphorylation data, together with the fact that expressed PR required hormone for DNA binding, indicate that the hormone-dependent phosphorylation step responsible for PR upshifts on sodium dodecyl sulfate-polyacrylamide gel electrophoresis is not required for receptor binding to DNA. The baculovirus expression system, therefore, may prove valuable in dissecting the functional role(s) for both hormone-dependent and hormone-independent PR phosphorylation.
Animals↗
Measurement of human chorionic gonadotropin during early pregnancy: a comparison of two immunoradiometric assays.
Evidence of implantation following either in vitro fertilization and embryo transfer (IVF-ET) or gamete intrafallopian transfer (GIFT) was obtained by collecting blood on days 10, 12, 14, 16, and 18 after oocyte recovery (day 0), and retrospectively measuring human chorionic gonadotropin (hCG). This was done using immunoradiometric assays for hCG, manufactured either by Serono Diagnostics Ltd. (MAIAclone) or Diagnostics Products Corporation (IRMA-count). The analysis of 63 serum samples by both kits showed a good correlation (r = 0.99) but the Serono (y) method gave values which were consistently greater (y = 1.58x + 4.89) than those of the DPC (x) method. A comparison of the hCG profile of singleton pregnancies measured with either the Serono (n = 33) or DPC (n = 22) kits gave a similar relationship. These results suggest that great care must be taken when comparing results from different laboratories using different kits. Also, consideration must be given to the possible loss of a continuous database should a laboratory change kit.
Chorionic Gonadotropin↗
Serum PP14 levels in a patient with Turner's syndrome pregnant after frozen embryo transfer.
Data are presented on serum levels of the pregnancy-associated placental protein (PP14) in an XO mother, pregnant after frozen embryo transfer. Steroid replacement before and during early pregnancy was provided with transdermal oestradiol and vaginal micronized progesterone. Characterization of a pre-treatment cycle indicated a physiological response by the endometrium. The pregnancy was associated with normal maternal blood levels of all hormones considered responsible for maintenance of early gestation but with consistently subnormal serum levels of PP14. Hitherto, PP14 has been believed to be an endometrial protein arising from the decidua during pregnancy, under the influence of ovarian steroids. The findings in this case suggest that other factors are involved in its production, possibly under the control of, or originating from, the maternal ovary.
Adult↗
Use of buserelin in an IVF programme for pituitary-ovarian suppression prior to ovarian stimulation with exogenous gonadotrophins.
Daily s.c. injections of buserelin were commenced in the mid-luteal phase of the preceding cycle in 118 women undergoing in-vitro fertilization (IVF) and embryo transfer. Ovarian and pituitary suppression was said to have been adequately achieved when serum oestradiol was less than 50 pg/ml, serum LH less than 2.0 IU/l, no ovarian cysts greater than or equal to 10 mm diameter were present and menstruation had occurred. Nine groups of women were retrospectively identified after the administration of buserelin for 12 days according to whether pituitary and ovarian suppression had been achieved or not, and the reason for extended buserelin treatment prior to ovarian stimulation. Upon adequate suppression, patients were grouped in terms of the duration of exposure to buserelin, and ovarian stimulation was then started by daily injections of human menopausal gonadotrophin. There appeared to be no differences in the ovarian response for women down-regulated by day 12, 19 or greater than or equal to 26 days; those women requiring extended buserelin treatment did equally well compared to those women down-regulating quickly, in terms of number of oocytes recovered and fertilization rate. Clinical pregnancy rates per embryo transfer were 27/68(40%), 8/33(26%) and 4/17(24%) for those women down-regulated by days 12, 19 or greater than or equal to 26 respectively, and were not significantly different.
Adult↗
Endocrine changes in women conceiving during treatment with an LHRH agonist.
We report on eight patients who conceived during pituitary desensitization with buserelin in the luteal phase of the menstrual cycle. Pregnancy was diagnosed between day 12 and 21 of buserelin administration. Analysis of serum luteinizing hormone on day 12 showed that pituitary desensitization occurred in conjunction with increasing production of ovarian steroid hormones. Serum concentrations of human chorionic gonadotrophin (HCG) were less than 10 IU/l on day 1 of buserelin administration for seven of the eight patients. The serum concentration of HCG on day 12 showed a median value of 722 IU/l (range 14.6-798 IU/l). Five of the eight patients were given HCG support (10,000 IU) following the diagnosis of pregnancy--three of these patients have ongoing pregnancies and the remaining two had blighted ova on scan. Of the remaining three patients, one had a singleton pregnancy which miscarried at 9 weeks, one had a blighted ovum on scan and bled per vagina shortly after this, and one bled per vagina prior to a scan being carried out. Our results show that pregnancy can occur during pituitary desensitization with buserelin, despite patients being counselled not to have unprotected intercourse in the cycle during which administration commences. An HCG assay on day 1 of buserelin administration is not helpful. Pregnancy should be suspected when ovarian steroid production persists despite complete pituitary down-regulation.
Adult↗
Fertilization in vitro of supernumerary oocytes following gamete intra-fallopian transfer (GIFT).
Gamete intra-Fallopian transfer (GIFT) was performed in 130 treatment cycles over a 17-month period. In 91% (118/130) of the cycles one or more oocytes were available for insemination in vitro and only GIFT cycles with supernumerary oocytes were included in the present study. Pituitary and ovarian suppression was achieved with buserelin followed by stimulation of multifollicular development by human menopausal gonadotrophin (HMG). Failure of supernumerary oocytes to fertilize was associated with a significantly reduced pregnancy rate (3/23; 13%) compared to cycles where fertilization occurred in vitro (35/95; 37%). These findings demonstrate that the outcome of IVF of supernumerary oocytes may be of particular diagnostic value in couples where the female partner has not conceived following treatment by GIFT after pituitary down-regulation with buserelin and ovarian stimulation with HMG.
Adult↗