A comparative analysis of second-trimester ultrasound dating formulae in pregnancies conceived with artificial reproductive techniques

A dataset of 64 pregnancies conceived by artificial reproductive techniques was studied to assess the accuracy of second-trimester dating formulae when these were applied in routine ultrasound clinics in different centers. Dating formulae for biparietal diameter (BPD) and femur length (FL) were derived for a gestational age range of 14-23 weeks. The best fit curves represented linear equations: gestational age (days) = 44.2 + 2 x BPD; and gestational age (days) = 67.4 + 2.3 x FL. Twelve published formulae for biparietal diameter and femur length were reviewed and systematic and random errors were calculated for these formulae when they were applied to second-trimester scan measurements in precisely dated pregnancies. Overall, published dating formulae performed well in predicting gestational age. The 95% confidence interval was 8.3 days for biparietal diameter and 10.2 days for femur length. The study confirms the accuracy of ultrasound dating in routine ultrasound clinics and supports the use of ultrasound measurement alone in preference to menstrual history for dating pregnancy.     

Partial and complete correlations of fetal maturity with ultrasonographic and enzymatic measurements

The computer-aided enzymatic and ultrasonographic method of pregnancy monitoring takes into account ultrasound measurements of fetal parameters (BPD, HC, AC, FL) and oxytocinase (CAP1) and isooxytocinase (CAP2) levels in maternal blood. In this paper we decided to verify which one of this values have the major influence on fetal maturity prediction. 385 pregnancies, after sterility treatment, were studied where ultrasound measurements and enzymes levels were estimated simultaneously as well as current fetal maturity were established using computer-aided method by M. Klimek. There are high sample correlations between current fetal maturity and the number of remaining days to labor (r = -0.8) as well as ultrasound measurements (BPD r = 0.87, HC r = 0.89, AC r = 0.90, FL r = 0.81). Fetal maturity correlations with enzymatic values (CAP1, CAP2) were smaller 0.55, 0.52 respectively. The partial correlation pictures this relations much stronger -0.29, 0.28, 0.22, 0.30, 0.46, -0.06, 0.13 respectively.     

Identifying patient needs in the context of the medication use situation

This study was designed to assess junctional zone contractions (JZ) during cycles of in-vitro fertilization (IVF) and embryo transfer in oocyte donors exposed to a long protocol regime for ovarian stimulation. Real-time transvaginal ultrasound and advanced audio-visual and computer technology were used to evaluate the contraction pattern, frequency and velocity. At the time of down-regulation JZ contractions were not observed. After 7 days of superovulation all patients displayed cervico-fundal, fundo-cervical and random contractions. Cervico-fundal waves dominated the picture at the time of human chorionic gonadotrophin injection. However, the activity was strongest on the day of oocyte retrieval. At that time the percentage of opposing waves increased and fundo-cervical waves disappeared. The highest wave frequency and velocity (4.29+/-0.68 waves/min and 2.73+/-0.54 mm/s respectively) were observed at the time of oocyte retrieval. All patients had some JZ activity on days 2, 3 and 4 after oocyte retrieval but regular wave-like contractility gradually decreased and only single random movements were seen on day 4 after oocyte retrieval. In conclusion, JZ activity throughout the IVF cycle is more exaggerated when compared to the results reported from observations of the natural cycle but follows a similar pattern. This fact can probably be explained by the vastly different hormone levels. Higher JZ activity and correspondingly increased mobility of the endometrium may impair its receptivity and affect implantation.     

Effect of routine screening for Down's syndrome on the significance of isolated fetal hydronephrosis

OBJECTIVE: To determine the risk of Down's syndrome in fetuses with isolated hydronephrosis at 18-23 weeks in an unselected general population after routine screening for Down's syndrome, using first trimester nuchal translucency measurement and second trimester maternal serum biochemistry. POPULATION: All pregnant women undergoing a routine 18-23 week ultrasound scan, from a population who had been offered screening for Down's syndrome. SETTING: A district general hospital serving a low risk obstetric population. METHODS: Prospective study of all routine 18-23 weeks ultrasound scans. The prevalence of isolated hydronephrosis and Down's syndrome was determined and the relative risk for Down's syndrome was calculated for different ultrasound findings. RESULTS: 10,971 women were scanned at 18-23 weeks during the study period. Down's syndrome was diagnosed in 14 of 20 cases before this stage using first trimester nuchal translucency measurement and second trimester maternal serum biochemistry. Isolated fetal hydronephrosis was diagnosed in 423 pregnancies (3.9%); none of these pregnancies were affected by Down's syndrome. The relative risk for Down's syndrome was 0.18 (95% CI 0.06-0.53) for women with a normal scan (n = 9983). When multiple ultrasound markers were found (n = 565), the relative risk for Down's syndrome was 2.00 (95% CI 0.18-22.10) and 9.00 (95% CI 1.14-71.30) for all other aneuploidies. CONCLUSION: The finding of isolated fetal hydronephrosis does not significantly increase the age-related risk for Down's syndrome. The presence of multiple ultrasound markers is associated with an increased risk of aneuploidies other than Down's syndiome. These findings are explained by the reduced prevalence of Down's syndrome as a result of prior screening and diagnosis of this condition.     

Endometrial stripe thickness as a predictor of ectopic pregnancy

We have examined the efficacy of highly purified follicle stimulating hormone (FSH-HP) for controlled ovarian stimulation in our in-vitro fertilization (IVF) programme, and compared the results obtained with this preparation with those using human menopausal gonadotrophin (HMG) in 15 patients who had received treatment with both FSH-HP and HMG in consecutive cycles (n = 39). No differences were found in the duration of stimulation, which was 13.9 days (HMG) as compared with 14.3 days (FSH-HP). However, in the FSH-HP-treated cycles we found a striking difference in the rise of serum oestradiol, which was significantly lower than in HMG-treated cycles (2953 +/- 938 pmol/l as compared with 6349 +/- 3683 pmol/l on the day before ovum retrieval). Number and size of follicles were similar in the two groups, as were oocyte characteristics. Increase in endometrial thickness at two days prior to ovum retrieval was slightly higher after HMG. The results indicate that in combination with a long gonadotrophin-releasing hormone agonist (GnRHa) protocol, pure FSH is sufficient for adequate follicle recruitment and growth. However, since FSH-HP resulted in markedly reduced concentrations of serum oestradiol as compared to HMG cycles, IVF programmes using repeated oestradiol measurements to decide the day of ovum retrieval must take this into consideration in order not to prolong the stimulation unnecessarily.     

Pregnancy and birth after high serum progesterone concentrations during the follicular phase in an in-vitro fertilization cycle with gonadotrophin-releasing hormone agonist suppression

This case illustrates the possibility of achieving a pregnancy and birth when elevated progesterone concentrations (> 4 ng/ml) are present during the follicular phase (from 6 days before human chorionic gonadotrophin injection) of a gonadotrophin-releasing hormone agonist/menotrophin cycle for in-vitro fertilization (IVF). The present patient underwent three IVF/embryo transfer cycles in which progesterone concentrations were repeatedly increased from the midfollicular phase onwards. A pregnancy was achieved after the first IVF attempt but ended in a miscarriage in the 19th week of gestation. During the second IVF attempt an endometrial biopsy taken on the day of oocyte retrieval revealed an endometrial advancement of 2 days. A successful pregnancy and birth was again achieved after the third IVF attempt although progesterone concentrations were considerably increased from 6 days before the ovulatory stimulus.     

Relationship of initial chorionic sac diameter to abortion and abortus karyotype based on new growth curves for the 16th to 49th post-ovulation day

In order to determine whether initial chorionic sac diameter is related to subsequent abortion, abortus karyotype, or birth weight and length, chorionic sac diameter was prospectively measured by transvaginal ultrasound in 700 singleton pregnancies before post-ovulation day 31, the latest day cardiac activity becomes detectable in normal pregnancy. Results were compared to values for the 10th to the 90th centiles, determined from 227 measurements of in-vitro fertilization and gamete intra-Fallopian transfer pregnancies. The abortion rate was 23.9% [95% confidence interval (CI) 19.2%, 28.6%] when initial chorionic sac diameter was below the 50th centile, compared to 6.9% (95% CI 4.9%, 9.4%) when equal to or above the 50th centile. Chorionic sac diameter was below the 50th centile in all anembryonic abortions and in 62% of embryonic abortions. Triploidy, trisomy 47 + 16, or trisomy 16 and the presence of satellite bodies on chromosome 22 were the only abortus karyotypes significantly associated with small chorionic sac diameter. Initial chorionic sac diameter was not associated with birth weight or length. We conclude that chorionic sac diameter is decreased in anembryonic and embryonic abortion and that normal pregnancy outcome may be expected in 90-95% of pregnancies in which initial chorionic sac diameter is equal to or above average.     

A new method for freezing testicular biopsy sperm: three pregnancies with sperm extracted from cryopreserved sections of seminiferous tubule

OBJECTIVE: To determine whether spermatozoa, located in the seminiferous tubules obtained by needle puncture testicular biopsy, could be cryopreserved successfully within the tubules and subsequently be used for in oocyte fertilization via intracytoplasmic sperm injection (ICSI) after the spermatozoa were removed from the thawed tubules. DESIGN: Clinical case series. SETTING: Private IVF unit. PATIENT(S): Six azoospermic patients (four obstructive, two maturation arrest). MAIN OUTCOME MEASURE(S): Survival rate of thawed spermatozoa, fertilization rate of oocytes after ICSI with spermatozoa extracted from thawed tubules and pregnancies. RESULT(S): All six patients had adequate motile spermatozoa extracted from the thawed tubule sections, and all patients achieved fertilization via ICSI with their partner's eggs. The fertilization rate was 46%, compared with 56% obtained in other previous patient cycles using fresh testicular spermatozoa. Three pregnancies resulted from five ETs. CONCLUSION(S): Cryopreservation and subsequent thawing of seminiferous tubules proved to be a simple and successful method for storage of testicular spermatozoa, reducing the need for repetitive testicular biopsies and increasing the likelihood of sperm availability on the day of oocyte retrieval.     

Serum pregnancy-specific beta1-glycoprotein before embryo transfer is related to endometrial thickness and to outcome prognosis in women undergoing in-vitro fertilization treatment

We have previously observed the repeated presence of low but detectable amounts of the trophoblast marker pregnancy-specific beta1-glycoprotein (SP1) in the serum of some women undergoing in-vitro fertilization (IVF) treatment around the time of oocyte retrieval. The occurrence of these signals seemed to be restricted to a defined group of patients which also showed a lower pregnancy success rate in a preliminary study. To test our hypothesis we have analysed 173 consecutive cycles leading to an embryo transfer. Fifty-four cycles (31%) had a serum SP1 level of at least 0.1 ng/ml between days embryo transfer -5 and embryo transfer (group A). Five pregnancies were obtained in this group (pregnancy rate = 9.3%), while in group B, defined by the absence of detectable SP1 before embryo transfer (119 cycles), 36 ongoing pregnancies were achieved (30.3%). Ten of the 41 pregnancies were achieved in 33 first-time non-pregnant patients undergoing further attempts during the study period. Again the pregnancy rate was higher in the first-time group B women (9/23 versus 1/10 for group A). Patients tended to remain in their groups A or B, the latter being associated with a better immediate as well as subsequent chance for pregnancy. Group A cycles had a significantly lower endometrial thickness two days before oocyte retrieval than group B (P = 0.0011). We postulate that the presence of an unknown, maternal and progesterone- or follicle stimulating hormone-independent factor in some patients could stimulate tonic ectopic SP1 synthesis and at the same time negatively influence endometrial development.     

Effects of fetal number and multifetal reduction on length of in vitro fertilization pregnancies

OBJECTIVE: To determine the effects of multifetal reduction and other variables on the duration of gestation of in vitro fertilization (IVF) pregnancies. METHODS: All 274 IVF pregnancies from the inception of the Women and Infants' Hospital IVF Program on May 26, 1988, until December 31, 1993, were evaluated. RESULTS: Spontaneous reduction occurred in ten pregnancies, and multifetal reduction was elected in 28 multiple gestations. Among 260 pregnancies that remained viable beyond 20 weeks, 162 singletons (37.9 +/- 0.29 weeks; mean +/- standard error) had a longer mean gestation than did 64 twins (34.6 +/- 0.61 weeks), 25 pregnancies reduced to twins (33.4 +/- 1.0 weeks), or nine triplets (29.7 +/- 1.9 weeks). Triplets delivered 4.9 weeks earlier than nonreduced twins (P < .05) and 3.7 weeks before twins resulting from multifetal pregnancy reduction (P < .05). Regression analysis showed that at the 8-week ultrasound, each viable fetus could be expected to reduce the duration of the gestation by about 3.6 weeks, and each fetus reduced medically or as a result of natural causes could be expected to prolong the gestation by approximately 3.0 weeks. Only 14% of triplet pregnancies underwent spontaneous multifetal reduction. CONCLUSION: Multifetal reduction of pregnancies with three or more fetuses was beneficial and increased the duration of gestation.     

Complications of vertebral metastasis surgery

The objective of the study was to evaluate the crown-rump length (CRL) in high-order multifetal pregnancies (three or more fetuses). The records of patients who underwent multifetal pregnancy reduction were reviewed. The following parameters were defined: the mean CRL (CRLMEAN) and the difference between the largest and smallest CRL (CRLRANGE) for each pregnancy. CRLMEAN was plotted versus gestational age (GA) and the line of best fit was derived. The residual for each CRL (CRLRES) was calculated by subtracting the value predicted by the regression line from the individual CRL (CRLIND). Regression lines for single-tons with confirmed GA from four previously reported studies were used for comparison. A total of 82 patients were included (29 triplets, 38 quadruplets and 15 quintuplets; mean GA 10.7 +/- 0.78 weeks). CRLMEAN correlated with GA (CRLMEAN = 38.88 - 8.78(GA) + 0.82(GA)2; R = 0.83; R2 = 0.70). This second-degree polynomial remained within the range for singletons. No significant differences in maternal age, GA and CRLMEAN were noted between the patients with different numbers of fetuses. The median CRLRANGE was largest in quintuplets, followed by quadruplets and triplets. The CRLRANGE correlated poorly with GA. The mean CRL in multifetal pregnancies increases with GA similarly to that in singletons. The variability of individual measurements increases with the number of fetuses and CRLs are lower in quintuplets.     

Human menopausal gonadotropin during in vitro maturation of human oocytes retrieved from small follicles enhances in vitro fertilization and cleavage rates

OBJECTIVES: To compare the IVF rates of oocytes retrieved from small follicles (< 2 mL in volume) with those of oocytes retrieved from large follicles and to test the effect of adding gonadotropins to the IVF medium on the fertilization rates of oocytes from small follicles. DESIGN: Oocytes were retrieved with endovaginal ultrasound (US) guidance from patients undergoing infertility treatment in our IVF program. Oocytes were grouped according to the volume of the originating follicle and subjected to our routine procedure for IVF. HMG was added to the IVF medium for some of the oocytes from small follicles. SETTING: Toronto Fertility and Sterility Institute is affiliated with the University of Western Ontario and University of Toronto and is equipped for RIA, endovaginal US monitoring and oocyte retrieval, and for processing and culturing gametes and embryos. PATIENTS: Infertile patients admitted to our IVF program. INTERVENTIONS: Patients underwent ovarian stimulation with hMG before oocyte retrieval. No other interventions were introduced to the processing and culturing the gametes and embryos except the addition of hMG to the medium of some of the small follicle-originated oocytes with the informed consent from the patients. MAIN OUTCOME MEASURES: Rates of fertilization, cleavage of the fertilized embryos before replacement, and meiotic status of some of the oocytes from small follicles. RESULTS: Most of the oocytes from small follicles did not complete the first meiotic division; they had low rates of fertilization and cleavage compared with oocytes from large follicles, and these rates were improved by the addition of hMG to the IVF medium. CONCLUSIONS: Oocytes from small follicles are probably less mature and require a more physiological environment to achieve normal rates of fertilization and cleavage.     

Histological evaluation of endometrium on the day of oocyte retrieval after gonadotrophin-releasing hormone agonist-follicle stimulating hormone ovulation induction for in-vitro fertilization

The objective of this study was to evaluate the histopathological characteristics of endometrial biopsies taken on the day of oocyte recovery in in-vitro fertilization (IVF) cycles with a satisfactory response to ovulation induction. A group of 33 patients who went through ovulation induction for IVF, and in whom an endometrial polyp was suspected on transvaginal ultrasonography during the monitoring phase, were studied. Following oocyte recovery, hysteroscopy, polypectomy and endometrial curettage were performed. Dating of endometrial glands and stroma was carried out in the tissue not containing the polyps. The total dose of follicle stimulating hormone (FSH), duration of ovulation induction, peak oestradiol and luteinizing hormone (LH) concentrations, thickness of endometrium and number of oocytes were recorded and compared to the endometrial dating of the specimens. In 15 cycles (45.5%), the endometrium was classified as 'in phase' (group I), 'advanced' by 2-4 days in a further 15 (45.5%, group II), and in the remaining three cycles (9%) it was delayed in maturation (group III). Younger age was correlated with advanced staging of the endometrium (r = -0.42; P = 0.015). Women with 'in phase' and 'advanced' maturation were similar in their response to ovulation induction; however, there was a strong correlation between advanced dating of endometrium and number of oocytes retrieved (r = 0.49; P = 0.04). Endometrial staging on the day of oocyte retrieval varied widely in patients treated by the same gonadotrophin-releasing hormone agonist (GnRHa)/FSH protocol for ovulation induction. This difference was not predictable by parameters monitored through the cycles.     

Outcome of pregnancy in IVF-ET cycle at Ramathibodi Hospital

As assisted reproductive technology is being developed, in vitro fertilization and embryo transfer (IVF-ET) are the treatments of choice for many infertility problems. The outcome of pregnancies achieved by IVF-ET is different from that of spontaneous pregnancies. In this retrospective study, the outcome of pregnancies from 400 IVF treatment cycles performed from October 1991 to October 1994 were reported. There were 80 pregnancies (20% per oocyte retrieval, 21.9% per embryo transfer) with an increased rate of abortion (30%), multiple pregnancy (20%), ectopic pregnancy (6.25%), heterotopic pregnancy (1.25%), preterm delivery (11.8%), low birth weight (35.8%) and cesarean section (62.7%). This study shows that the complication rate of pregnancies from IVF-ET cycles was higher than that found in spontaneous pregnancies. Some complications such as multiple pregnancies may be prevented by limiting the number of transferred embryos. From this study, all IVF-ET pregnancy should be considered as high risk pregnancies.     

Regeneration of defects in articular cartilage with callus and cortical bone grafts. An experimental study

Luteal-phase estrogen and progesterone concentrations were measured every other day and used to monitor the corpus luteum activity. The patterns of estrogen and progesterone concentrations were compared relative to the day of endogenous human chorionic gonadotropin (hCG) detection (defined as the day of implantation). The relationship between estrogen and progesterone and hCG concentrations was studied in 71 viable pregnancies, 12 clinical abortions, five preclinical abortions and 84 non-pregnant cycles after IVF/ET. Although all patients received luteal-phase progesterone support (25-50 mg/ml), low late luteal-phase progesterone concentrations of < 30 ng/ml from day + 11 to day + 15 were found in 64 patients (17% of viable pregnancies, 33.3% of clinical abortions, 60% of preclinical abortions and 53.6% of non-pregnant cycles) day + 1 was the day of retrieval). Implantation always occurred before or on day + 13 and 86% of pregnant cycles implanted on day + 8 to day + 11. Viable pregnancies had significantly higher mean progesterone concentrations on day + 3 to day + 7 (pre-implantation) and on day + 9 to day + 15 (postimplantation) than those of non-pregnant cycles or abortions. On the day of implantation, the mean +/- standard of deviation of estrogen (pg/ml) and progesterone (ng/ml) levels for viable pregnancies, clinical abortion and preclinical abortions were 314 +/- 210, 40.5 +/- 25; 226.7 +/- 246, 48.7 +/- 31; and 39.6 +/- 24.5, 28.6 +/- 24.5, respectively. On the same day, 73.2% of viable pregnancies, 41.7% of clinical abortions, and 20% preclinical abortions had a progesterone concentration > 30 ng/ml; 73.2% of viable pregnancies, 41.7% of clinical abortions and 20% of preclinical abortions had an estrogen concentration > 100 pg/ml. Although not precluding implantation completely, late luteal-phase hormonal deficiencies may impair endometrial growth and might ultimately lead to failure or abnormal implantation. A viable pregnancy requires not only a functional corpus luteum in the early luteal phase to develop a receptive endometrium, but also a responsive corpus luteum in the late luteal phase to support pregnancy. The time of implantation is critical. Implantation that occurs before the demise of the corpus luteum will facilitate a normal pregnancy.     

Psychological reactions during in-vitro fertilization: similar response pattern in husbands and wives

The purpose of this study was to examine differences in daily emotional, physical and social reactions among husbands and wives during in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Forty couples about to undergo ICSI or IVF at a private infertility clinic monitored their emotional, physical and social reactions daily for one complete treatment cycle from the first day of stimulation until the outcome of treatment was known (approximately 35 days). The results showed that men and women had a similar response pattern to oocyte retrieval, fertilization, embryo transfer and the pregnancy test. These stages were associated with the most significant changes in reactions for both spouses. The pattern of results suggested that the most important psychological determinant of reactions during IVF was the uncertainty of treatment procedures. Spouses appeared to be equally sensitive to this uncertainty and both appeared to respond to it with ambivalent feelings involving emotional distress and positive feelings of hope and intimacy.     

Color Doppler ultrasound velocimetry of internal carotid arteries in normal pregnancies

BACKGROUND: There are few reports on maternal cerebral circulation during pregnancy. Using the hypothesis that cerebral blood flow resistance decreases progressively with advancing gestational age (GA) as a consequence of estrogenic effects on the vascular system during pregnancy, we calculated the pulsatility index (PI) and the mean velocity (MV) of the maternal right internal carotid artery (ICA) in order to build fitted reference centiles. METHODS: A total of 315 pregnant women with a single fetus were studied at 20-42 weeks' gestation. The patients had uncomplicated singleton pregnancies and were without any known fetal or maternal risk factors. Duplex color ultrasound with a pulsed Doppler scanner (7.5 MHz) was used to determine the PI and MV of the maternal ICA. RESULTS: Among the 315 healthy pregnant women, the PI of maternal ICA decreased progressively with advancing GA, PI50th = Exp (0.3124-0.0084 x GA), (p = 0.0003), during the second half of pregnancy. The predicted 50th percentile PI values of the ICA decreased from 1.117 (5th% = 0.755, 95th% = 1.654) at 24 weeks' gestation to 0.917 (5th% = 0.659, 95th% = 1.448) at 40 weeks' gestation. The MV of the maternal ICA also decreased progressively with advancing GA, MV50th = Exp (3.855-0.0093 x GA), (p = 0.0005), during the second half of pregnancy. The predicted 50th percentile MV values in the ICA decreased from 37.811 cm/sec (5th% = 24.177 cm/sec, 95th% = 59.138 cm/sec) at 24 weeks' gestation to 32.591 cm/sec (5th% = 20.818 cm/sec, 95th% = 51.021 cm/sec) at 40 weeks' gestation. CONCLUSIONS: Both the PI and MV of the maternal ICA decreased with GA during the second half of normal pregnancy as a result of intracerebral vessel dilatation.     

Daily measurements and in-vitro effects of human chorionic gonadotrophin in the early luteal phase

The purpose of the present study was to analyse daily measurements of human chorionic gonadotrophin (HCG) in in-vitro fertilization (IVF) cycles and to reproduce the effects of HCG in vitro using human granulosa-luteinized cells from the same patients. The study population consisted of nine women undergoing IVF because of tubal infertility in whom blood was drawn every 24 h from the day of the ovulatory dose of HCG (10,000 IU) until 6 days after ovum pick-up. Granulosa-luteal cells from the follicular aspirates were collected and cultured in vitro up to 6 days in the presence of increasing concentrations (0, 0.01, 0.1, 1.0 and 100.0 IU/ml) of HCG. Serum progesterone and HCG in vivo as well as progesterone accumulation in vitro on days 2, 4 and 6, were the main outcome measures. Maximum HCG concentrations (0.25 IU/ml) were reached the day before ovum pick-up, and continuously decreased until day 6 after ovum retrieval. HCG did not stimulate progesterone production in vitro at any dose tested until day 6 after ovum pick-up. Then, 0.01 IU/ml resulted significantly (P < 0.05) stimulatory compared to controls, while 1.0 IU/ml was inhibitory (P < 0.05). It is concluded that HCG supplementation in an IVF cycle is unnecessary until day 6 after ovum pick-up. On day 6, progesterone production is stimulated with very low concentrations of HCG.     

A new source of human oocytes: preliminary report on the identification and maturation of human preantral follicles from follicular aspirates

This study aimed to investigate the development of human preantral follicles and oocyte maturation in vitro. Preantral follicles were obtained from follicular aspirates during egg retrieval carried out during an in-vitro fertilization (IVF) programme. They were first incubated in Ham's F10 medium with 15% fetal cord serum (FCS). After 28 days, the medium was supplemented with different doses of human menopausal gonadotrophin (HMG), human follicular fluid (hFF) and epidermal growth factor (EGF) by orthogonal design. Promotion of final maturation was completed in the presence of HMG and hFF. Development from preantral to antral follicles was found within 6-12 days of culture. With time, the proportion of follicles with diameters of >300 microm increased at 21-28 days of culture (P < 0.005). The maximum number of oocytes extruded, and first polar body formation, occurred in the presence of 0.15 IU/ml HMG 40% (v/v) hFF and 6 ng/ml EGF. We conclude that follicular aspirates obtained during egg retrieval in an IVF programme contain many preantral follicles which could develop into antral follicles with extrusion of oocytes in culture, and that the oocytes can mature in vitro. Hence, a new source of human oocytes is available.