Comparison of two culture systems for the in-vitro growth and maturation of mouse preantral follicles.

The objective of our study was to compare to ability of collagen-treated membranes and bovine collagen gels to maintain murine preantral follicle growth and development in-vitro. To fulfill that objective, murine follicle and oocyte growth rates were followed for ten days in culture. Meiotic competence and the capacity to reach the two-cell stage after in-vitro maturation and fertilization, respectively, were then assessed. We used preantral follicles from 12 day-old CF-1 female mice that were isolated by enzymatic digestion from ovaries. Follicles were placed either on collagen-treated membranes or embedded in a bovine collagen matrix. The follicles were grown, changing the media and obtaining measurements every other day for ten days. Following culture, the granulosa-oocyte complexes were matured; the resultant metaphase II arrested oocytes were inseminated and cultured to the two-cell stage. The data was analyzed with significance considered for probability values of p < 0.05. We performed individual measurements on 650 follicles in seven separate experiments. Forty-eight hours after initial seeding and throughout the entire length of culture, both the follicles and oocytes grown in the collagen matrix were larger than follicles cultured on collagen-treated membranes (p < .0001). However, oocyte recovery rates were higher among follicles cultured on collagen-treated membranes (p < .01). Similar percentages of meiotically competent oocytes, fertilization and cleavage rates were observed in both groups. Our results show that mouse preantral follicles display a greater growth rate when grown embedded in a collagen gel matrix. This may be due to the maintenance of a normal three-dimensional organization of the follicle within the collagen matrix. However, this system does not enhance meiotic competency or fertilization rates in the mouse when compared to culture on collagen-treated membranes.

Timing of the endometrial biopsy may be critical for the accurate diagnosis of luteal phase deficiency.

OBJECTIVE: To determine the optimal time to perform the endometrial biopsy for the detection of "out-of-phase" endometrium. DESIGN: Two endometrial biopsies were performed during a single menstrual cycle in each subject. The patient's chronological day was determined by counting forward from the midcycle LH surge, as assessed by urinary LH detection. The "early" biopsy was done on day LH + 7.4 +/- 0.8, and the "late" biopsy on day LH + 11.6 +/- 0.7. Each biopsy was independently read by two pathologists and was considered out of phase if the histologic date was > or = 3 days delayed compared with the chronological date. SETTING: Infertility practice of an academic teaching hospital. PATIENTS: Thirty-three ovulatory women seeking evaluation for infertility. MAIN OUTCOME MEASURE: Number of patients with out-of-phase endometrium detected by the early versus the late biopsy. RESULTS: There was a significantly greater detection rate for out-of-phase endometrium using the early biopsy (12.1% to 18.2% incidence depending on the observer) compared with the later biopsy (6.1% to 9.1% incidence). A majority of the early out-of-phase biopsies corrected by the time of the later biopsy. CONCLUSION: Our findings indicate that an endometrial biopsy performed in the midluteal phase may detect a greater number of women with delayed endometrial maturation during the temporal window of embryo implantation. The observation that most of the women with out-of-phase midluteal biopsies had normal late luteal endometrium may represent a cryptic form of luteal phase deficiency.

Embryonic heart rate in human pregnancy.

Seventy-two women with known gestational ages underwent serial human chorionic gonadotropin (hCG) measurements and transvaginal ultrasound studies with embryonic heart rate measurements. In 53 continuing singleton pregnancies, embryonic pulse appeared between days 26 and 32, was 80 beat per minute (bpm) on day 26, and increased linearly to plateau at 160-200 bpm by day 45 (r2 = 0.72). The pulse was always seen with hCG greater than 21,000 mIU/mL; pulse rate was correlated to embryonic crown-rump length. Of the remaining 19 pregnancies, 8 were anembryonic, 10 showed heart activity but subsequently aborted, and 1 was terminated. Absence of embryonic pulse by 32 days after conception or a serum hCG greater than 21,000 mIU/mL predicts spontaneous abortion; presence of a pulse may not guarantee successful continuation of the pregnancy since incidence of spontaneous abortion after visualization of embryonic pulse may be as high as 16%.