SVIBOR - Project code: 3-01-089


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SVIBOR - Collecting Data on Projects in Croatia

Project code: 3-01-089


Main researcher: SUCHANEK, ERNEST (98740)

Type of research: applied
Duration from: 04/30/91. to 12/31/95.

Papers on project (total): 11
Papers on project quoted in Current Contents: 5
Institution name: Medicinski fakultet, Zagreb (108)
Department/Institute: Department of Clinical Biochemistry and Reproductive Biology University Clinic of Obstetrics and Gynecology
Address: Petrova 13
City: 10000 - Zagreb, Croatia
Phone: 385 (0)1 44 40 22
Fax: 385 (0)1 44 42 79
Phone: 385 (0)

Summary: To determine the concentrations of hyaluronic acid,FSH, progesterone and estradiol in the follicular fluid obtained from in vitro fertilization-embryo transfer (IVF-ET) patients and to assess the value of these measurements in predicting the outcome of fertilization. One hundred and eleven samples were retrospectively analyzed for the hyaluronic acid and hormone contents. Preovulatory follicular fluid samples were collected from 67 women undergoing IVF-ET treatment due to tubal absence or obstruction. The follicular fluid hyaluronic acid and hormone concentrations were compared according to the type of ovulation induction, follicular development, and IVF outcome. According to the type of ovulation induction, a significantly lower hyaluronic acid concentration was found in follicular fluid harvested from the patients treated with GnRH-agonist-human menopausal gonadotropins. No significant correlation was found between follicular fluid hyaluronic acid and either morphologic maturity of the oocyte-cumulus complex or fertilizability of oocytes. The level of FSH was significantly higher in follicular fluids yielding a mature oocyte-cumulus complex, and from which the oocyte obtained successfully fertilized and cleaved. A significant increase in the estradiol concentration was found in follicular fluid in which mature cumuli oophori were present. The levels of hyaluronic acid significantly correlated with FSH in follicular fluid. Expansion of the human oocyte-cumulus cell complex is a FSH-dependent phenomenon. The data are in agreement with the hypothesis that intrafollicular FSH plays an important role in the secretion of hyaluronic acid by granulosa cells and may act synergistically with estradiol to enhance cytoplasmic maturation resulting in successful fertilization.Preovulatory serum estradiol and progesterone levels as well as their ratio were compared in different types of ovulation induction in order to determine whether these findings could be used to predict the number of preovulatory follicles, number of oocytes aspirated and embryos transferred. Significantly more oocytes were obtained by follicular aspiration and significantly more embryos transferred to the uterus in patients receiving GnRH-agonist and human menopausal gonadotropins than in those given other ovulation inductors. On days -2 and -1 of the cycle, serum estradiol levels were significantly lower in pure FSH induction. Serum progesterone was significantly higher in pure FSH cycles on days -4 and -3. In clomiphene citrate and human menopausal gonadotropin induction, progesterone levels were significantly lower on days -2 and -1, and on the day of follicular aspiration. Ratios of estradiol/ progesterone were lower in pure FSH group from day -3 to day -1 of the cycle. A significant correlation was found between estradiol and progesterone serum levels and the numbers of preovulatory follicles, oocytes and embryos as well. The study revealed the usefulness of serum estradiol and progesterone determinations in assisted reproduction. Investigation was performed on estradiol and progesterone syntheses from granulosa cell culture concerning the composition of media used in oocyte and embryo culture. Menezo B2 and MEM media with the supplementation of 7% and 15% of newborn cord serum were used. Ovulation induction with human menopausal gonadotropins was performed in 11 women. From the 50 000 granulosa cells isolated the production of estradiol and progesterone during 4 days of culture was determined. The estradiol concentration was significantly higher in both media when 15% cord serum was used. Comparing the media with the same amount of cord serum no difference in estradiol production was noted. Higher progesterone synthesis was found in Menezo B2 with 7% than 15% cord serum supplementation. Progesterone was lower from the culture of granulosa cells in MEM than Menezo B2 with the same amount of cord serum. The results revealed the data that the composition of medium used for human oocyte and embryo culture is very important in the view of the success of assisted reproduction.

Keywords: Follicular fluid, hyaluronic acid, FSH, estradiol, progesterone, ovarian induction, in vitro fertilization, menstrual cycle, clomiphene citrate, human menopausal gonadotropins, gonadotropin-releasing hormone agonist, granulosa cell culture

Research goals: The aim of this study is to determine biochemical markers in serum, preovulatory follicular fluid and granulosa cells culture as well, to obtain data about oocyte maturity and their ability to fertilize in vitro. The results will be used for the type of pharmacological ovulation induction management and for endocrine environment composition which is of great importance for ovarian follicle physiology.


  1. Name of institution: Department of Obstetrics and Gynecology, Northwestern University Medical School and Andrology Laboratory Services, Inc.
    Type of institution: University/Faculty
    Type of cooperation: Joint publishing of scientific papers
    City: 60611 - Chicago, Illinois, S.A.D.

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Last update: 10/05/95