Developmental potential of microbiopsied murine blastocysts.

OBJECTIVE: To evaluate the frequency of mouse pups born following blastocyst biopsy and embryo transfer compared to nonbiopsied controls. METHODS: ICR Swiss albino and C57B1/6 mice served as embryo donors. Pregnant mare serum gonadotropin treatment was followed 46-52 hours later by hCG, when donors were paired with fertile males. Mating was confirmed the following day and embryos were collected on the afternoon of day 4. After overnight culture, hatching trophoblast was excised by micromanipulation with a fine-pulled glass pipette. Embryos (206 controls, 206 biopsied) were transferred to 26 pseudopregnant recipients. Alternate mouse strains were used to identify pups born from control or biopsied embryos. RESULTS: The end point was percentage of pups born ((number born/number transferred) x 100), using angular transformation before analysis. There was no significant difference (P > .1) between percent live-born in control (27.7%) or biopsied (34.5%) embryos, nor were there any strain differences. CONCLUSION: These data support the hypothesis that the developmental potential of murine blastocysts is not affected adversely by the biopsy procedure.

Growth of mouse pups derived from biopsied blastocysts.

OBJECTIVE: To test the hypothesis that removal of cells from the pre-embryo affects intrauterine growth, as reflected in birth weight or growth to weaning. METHODS: ICR Swiss albino and C57Bl/6 mice served as embryo donors to provide two strains of distinct coat colors. Alternate strains were assigned to control treatment or blastocyst biopsy of trophectoderm. Embryos from both treatments were transferred to a different uterine horn within each pseudopregnant recipient. A total of 412 embryos (206 controls and 206 biopsied blastocysts) were transferred to 26 recipients, which resulted in the birth of 122 pups. RESULTS: There was no significant difference (P > .1) in birth weight or growth to weaning, defined as [[(weaning weight-birth weight)/birth weight)] x 100], between male and female pups. There was a significant difference (P < .01) between pups derived from ICR (birth weight 1.9 +/- 0.02 g; growth to weaning 544 +/- 14%; n = 72) versus C57Bl/6 (birth weight 1.43 +/- 0.03 g; growth to weaning 460 +/- 14%; n = 50) embryos. However, there was no significant difference (P > .1) in birth weight or growth to weaning for pups derived from biopsied embryos (birth weight 1.70 +/- 0.03 g; growth to weaning 502 +/- 14%; mean +/- standard error of the mean; n = 67) or control embryos (birth weight 1.71 +/- 0.05 g; growth to weaning 520 +/- 17%; n = 55). CONCLUSIONS: These data support the null hypothesis that embryo biopsy does not result in reduced birth weight or altered subsequent growth to weaning.

Failure to demonstrate significant antisperm antibodies in peritoneal fluid of patients with endometriosis.

Endometriosis, even in mild cases, decreases monthly fecundity. Immunologic disorders have been suggested as the mechanism. In light of possible increases in serum autoimmune antibodies, increased peritoneal macrophages, and increased sperm phagocytosis associated with this disease, we postulated that peritoneal fluid antisperm antibodies would be increased and might be the cause of increased sperm phagocytosis and its associated infertility. Peritoneal fluid, from 18 patients with endometriosis and 10 infertile controls, was tested with the antisperm antibody immunobead test validated for peritoneal fluid. One of 18 patients with endometriosis and none of 10 controls had antisperm antibodies present. Therefore, increased sperm phagocytosis is unlikely a result of peritoneal antisperm antibodies in endometriosis patients.

Use of endometrial measurement as an exclusion criterion for in vitro fertilization using clomiphene citrate.

OBJECTIVE: Previous reports have indicated an association between endometrial development and pregnancy outcome for patients treated with clomiphene citrate (CC) in conjunction with intrauterine insemination or intercourse. We expanded the use of CC for ovulation induction in association with in vitro fertilization (IVF). This study was designed to determine if endometrial thickness should be used as an inclusion or exclusion criterion for CC-IVF. STUDY DESIGN: One hundred twenty-eight patients were enrolled in an ovulation-induction regimen using CC for expected IVF-ET between January 1992 and December 1992. A total of 81 patients met inclusion criteria for CC-IVF and had endometrial measurement performed prior to human chorionic gonadotropin administration. Patients were categorized on the basis of endometrial measurement as follows: (A) > 4 - < 7 mm, (B) > or = 7 - < or = 10 mm, and (C) > 10 mm. Standard IVF was performed, and pregnancy rates for each category were evaluated. RESULTS: A total of 23 pregnancies (28% per retrieval) were established. Pregnancy rates were not different by category (P > .10, Fisher's Exact Test): (A) 3/15 (20%), (B) 13/41 (32%), and (C) 7/25 (28%). CONCLUSION: These data suggest that for CC-IVF. endometrial measurement should not be used as an exclusion criterion since pregnancies occurred at comparable frequencies in all the groups.

Single injection of methotrexate for treatment of ectopic pregnancies.

OBJECTIVE: Our purpose was to evaluate the efficacy of single-dose methotrexate as a treatment option for ectopic pregnancies. STUDY DESIGN: Patients were recruited from a tertiary teaching hospital setting. They had (1) a gestational sac < 3.5 cm, (2) no fetal cardiac activity, and (3) no significant pelvic pain or signs consistent with hemoperitoneum. Patients excluded were those who did not desire future fertility or who had evidence of renal or liver disease. Sixty-one patients meeting these criteria were selected and treated with a single intramuscular injection of methotrexate at a dosage of 50 mg/m2. RESULTS: Of the 61 patients treated, 16 required a second injection and nine required surgical intervention. Fifty-two (85%) were successfully treated as outpatients with methotrexate alone. CONCLUSION: Nonsurgical treatment of ectopic pregnancies with single-dose methotrexate is an option for some patients, but more studies are needed to establish the safety, efficacy, and effect on fertility.

Feasibility of semen collection and cryopreservation during chemotherapy.

Collection and cryopreservation of semen was undertaken in a 40-year-old man undergoing combination cyclophosphamide-doxorubicin-vincristine-prednisone chemotherapy for diffuse large cell lymphoma. Semen samples worthy of cryopreservation could be obtained through 39 days of therapy, with azoospermia observed after 70 days. Although pregnancy has not been attempted in the current case, reports by others suggest that normal outcomes can be expected with thawed semen collected during chemotherapy.

Trophectoderm microbiopsy in murine blastocysts: comparison of four methods.

PURPOSE: This study compares four trophectoderm microbiopsy techniques for removal of blastomeres from murine blastocysts: (1) aspiration, trophectoderm pipetted through the zona; (2) incision, trophectoderm excised with a microrazor; (3) slit/excision, the zona slit and herniating trophectoderm excised; and (4) hatch/excision, trophectoderm cells excised after spontaneous hatching. RESULTS: Murine blastocysts were comparatively biopsied using one of four methods and contrasted to zona slit and nonmicromanipulated controls. Operative cellular injury was assessed by uptake of fluorescein isothiocyanate (FITC). Postoperative embryonic viability was assessed by blastocoele reexpansion and hatching inner cell mass development and trophectoderm plating. All techniques yielded cells available for genetic analysis. CONCLUSIONS: The slit/excision technique and hatch/excision techniques exhibited lower operative injury and the higher postoperative viability than aspiration or incision. The slit/excision and the hatch/excision techniques, though requiring two operative steps, appear to be the least damaging of the four methods. Therefore, they should be most applicable to human blastocysts obtained either by extended culture in vitro or by uterine lavage.

Predictive value of hCG level 14 days after embryo transfer.

OBJECTIVE: It has been reported that the quantitative serum hCG level 14 days after embryo transfer (ET) correlated with pregnancy outcome as well as a likelihood of a multiple gestation pregnancy. This prospective study was designed to assess the predictive value of a 14-day post-ET hCG level with pregnancy outcome and multiple gestation pregnancies. METHODS: Patients undergoing in vitro fertilization (IVF) and ET were monitored by serum quantitative hCG levels 14 days after ET. If positive, serial values of hCG were obtained and transvaginal ultrasound was performed 3 weeks after ET and weekly until fetal cardiac activity was seen. Ongoing pregnancies were defined as greater than 20 weeks. RESULTS: One hundred eleven patients had positive serum quantitative hCG levels 14 days post-ET; 89/111, or 80.2%, had ongoing pregnancies. The spontaneous miscarriage rate was, therefore, 19.8% (22/111). If the level was less than 300, the ongoing multiple pregnancy rate was 9% (5/57). If the level was between 300 and 600, the ongoing pregnancy rate was 40% (10/25). If the hCG level was greater than 600, the multiple pregnancy rate was 100% (7/7). CONCLUSIONS: These data support the hypothesis that hCG levels greater than 200 mIU/ml on 14 days post-ET are more likely to have ongoing pregnancies; hCG levels greater than 600 have a high likelihood of a multiple gestation pregnancy.