Ilan Calderon

A simplified coculture system with luteinized granulosa cells improves embryo quality and implantation rates: a controlled study.

OBJECTIVE To investigate the use of a simplified short-term coculture system with luteinized granulosa cells (GCs) in patients with failed IVF-ET. DESIGN Controlled clinical study. SETTING IVF unit, Department of Obstetrics and Gynecology, Carmel Medical Center. PATIENT(S) Patients with poor embryo quality in their previous IVF-ET cycles. INTERVENTION(S) Embryos from 40 patients, in which > 50% of the embryos were classified as poor quality in their previous IVF attempts, were grown on autologue GC culture system for a short period (24-48 hours) before being replaced in the uterine cavity. MAIN OUTCOME MEASURE(S) Embryo quality. RESULT(S) Significant decrease in poor quality embryos and increase in the proportion of good quality embryos were observed using a coculture system with autologue human GCs. Pregnancy rates in this groups of patients reached our standard IVF results during the same period. CONCLUSION(S) This study describes a simplified short-term coculture system with human autologue GCs. Poor quality embryos may be rescued to cleave regularly.

Treatment of hirsutism with cimetidine: a prospective randomized controlled trial *

Cimetidine (Tagamet, Smith Klein, Brussels, Belgium) the histamine receptor type 2 blocker, has been suggested as a possible treatment of hirsutism. In a prospective randomized controlled trial, cimetidine 1.5 gm a day was given for 3 months to 12 moderately or severely hirsute women, while 8 other women were randomized to serve as a control group. There was no significant change in the hair growth rate in either group as measured by two assessment methods. This finding suggests that cimetidine, given in the recommended dose for a period of 3 months, does not have a beneficial effect on hirsutism.

Pure follicle-stimulating hormone as an adjuvant therapy for selected cases in male infertility during in-vitro fertilization is beneficial

OBJECTIVES Since research has demonstrated the possibility of hormonal therapy for male infertility, we conducted a study to analyze the efficacy of pure follicle-stimulating hormone (pFSH) treatment in patients with idiopathic, severe oligoteratoastheno-spermia (OTA) syndrome, or failed fertilization before referral to an intracytoplasmic sperm injection (ICSI) in an in-vitro fertilization (IVF) program. STUDY DESIGN A retrospective, clinical study was carried out on 178 men with OTA syndrome. Group I comprised 76 patients selected for treatment with pFSH. Group II comprised 102 men who served as the controls. Pure FSH was administered intramuscularly to the patients in group I. Upon cessation of therapy, an IVF treatment cycle was carried out. RESULTS After treatment with FSH, sperm motility was the only parameter which significantly improved in Group I (34% vs. 23%, and 15% vs. 24% in the subgroups of Group 1, respectively; P<0.05). In-vitro fertilization pregnancy rates were similar in both groups. The highest spontaneous pregnancy rates were achieved in FSH-treated, severe OTA patients. CONCLUSIONS Patients with severe male factor infertility may benefit from pFSH in terms of sperm motility, fertilization by IVF and spontaneous pregnancy rates. Selection criteria for FSH treatment are proposed.

Sphingosine-1-Phosphate may Decrease the Cytotoxic Effect of Doxorubicin on Human Granulosa Cells

Context: The increase in malignancy of young women in the recent decades, combined with a significant improvement in long term survival after gonadotoxic chemotherapy, have brought about a ubiquitous interest in preservation of fertility in these young patients. The present study examines the effects of Sphingosine-1-Phosphate (S1P) on primary human granulosa cell cultures in-vitro as a possible protecting factor against Doxorubicin (DOX) and Cyclophosphamide associated toxicity. Understanding cytotoxic effects and gonadotoxicity in human luteinized Granulosa Cells (GC) may contribute to our understanding and preventing follicle loss. Study objective: To examine the possible protective effect of S1P on chemotherapy induced gonadotoxicity, in human luteinized Granulosa Cells (GCs). Design: Human GC’s were donated by women undergoing follicular aspiration for in vitro fertilization (IVF), after informed consent and institutional approval by ethics committee (IRB, Helsinki). The GCs were separated from RBC’s by centrifugation on ficoll and plated on multiwell plates for Lactate Dehydrogenase (LDH) assay, and on 6 well plates for flow cytometry. Each experiment was conducted in triplicates and repeated at least three times. Results: S1P significantly protected GCs against Doxorubicin (DOX) toxicity, but inconsistently against Cyclophosphamide. Conclusion: S1P may minimize the gonadotoxic effect of chemotherapy on human luteinized granulosa cells.

Environment of the preimplantation human embryo in vivo: metabolite analysis of oviduct and uterine fluids and metabolism of cumulus cells**Supported by IVF America Inc., Greenwich, Connecticut and Monash IVF Pty. Ltd., Melbourne, Victoria, Australia.

OBJECTIVE To determine the levels of metabolites surrounding the human oocyte and embryo in vivo. DESIGN Oviduct and uterine fluids were collected throughout the menstrual cycle. Cumulus cells were collected at oocyte retrieval and their production of metabolites was assessed. Samples were analyzed for pyruvate, lactate, and glucose by microfluorimetry. PATIENTS Luminal fluids were collected from naturally cycling patients at the time of routine clinical investigation. Patient consent and hospital ethics approval were obtained for this study. RESULTS Pyruvate in the oviduct did not vary with the day of cycle, the mean value was 0.24 mM. Lactate and glucose concentrations varied with the day of cycle; lactate increasing from 4.87 mM in the follicular phase to 10.50 mM at the time of ovulation, whereas glucose decreased from 3.11 mM in the follicular phase to 0.50 mM midcycle and subsequently increased to 2.32 mM in the luteal phase. The concentrations of pyruvate, lactate, and glucose in uterine fluid remained constant throughout the cycle (0.10, 5.87, and 3.15 mM, respectively). All metabolite concentrations in uterine fluid were significantly different from those in the oviduct midcycle. Cumulus cells readily consumed glucose in vitro, with lactate being the major metabolite produced. CONCLUSION These data indicate that lactate and glucose concentrations in the oviduct change with day of cycle and that the human embryo is exposed to different metabolite concentrations as it passes along the tract. Furthermore, cumulus cells readily consume glucose, producing lactate. Therefore, the early human embryo is exposed to low glucose and high lactate levels in vivo.