P-183
cantly higher in both eutopic and ectopic endometrial tissues of women
HUMAN OVARIAN BIOPSIES AS A VIABLE SOURCE OF PREAN-
TRAL FOLLICLES. S. Rice, K. Ojha, H. D. Mason. Basic Medical Sci-
ences, St Georges, University of London, London, United Kingdom;
Clinical Developmental Sciences, St Georges, University of London,
London, United Kingdom.
OBJECTIVE: The study of human preantral follicles has been restricted
by the limited amount of tissue available. Our aim was to determine the utility
of ovarian biopsy for follicle research and IVM by assessing the retrieval
rates and developmental stage of preantral follicles from this tissue and the
amount of cDNA obtained from these follicles.
DESIGN: Follicle isolation from ovarian tissue.
MATERIALS AND METHODS: Ovarian cortical biopsies were obtained
from St Georges Hospital, London with ethical approval and informed con-
sent from women undergoing C-sections (n 12, CB) or TAH/BSO (n 7,
OV) for various benign gynaecological conditions. Follicles were manually
dissected from ovarian stroma after a brief enzymatic digestion, photo-
graphed and the developmental stage and morphology of each follicle ascer-
tained. Follicles were lysed, RNA extracted, reverse transcribed to cDNA and
the quantity of cDNA measured.
RESULTS: The mean age was 36.9 yrs and was similar in both groups. A
total of 351 follicles was retrieved and 249 were classifiable. All samples
yielded follicles except for two from women aged over 40 yrs. The average
yield was 13.1 follicles/patient with more follicles/patient retrieved from CB
samples (22.3) compared to OV samples (8.4). There were 80 primordial, 53
transitional, 82 primary, 26 secondary and 8 multi-laminar follicles. 71 fol-
licles were retrieved from <30 yr group (n 3), 167 from 3139 yr group
(n 11) and 11 from the 40 yr group (n 5). There was an age-related Peoples Republic of Korea.
decline in the mean follicle numbers/patient from 23.7  1.7 (<30 yr)
down to 15.2  3.1(3139 yr) and 2.2  0.95(40 yr) (mean  SEM,
r2 0.986). Overall more follicles at all stages were obtained per patient
from the <30 yr compared to the other groups (ANOVA P0.01). The
mean amount of cDNA obtained was 1.17  0.05 mg/ml (n 11 primordial),
1.38  0.09 (n 10 transitional),1.33  0.06 (n 11, primary), 1.42  0.09
(n 9, secondary) (mean  SEM).
CONCLUSIONS: In spite of irregular distribution of follicles throughout
the ovary, cortical samples provide a useful source of preantral follicles. The
sensitivity of genomic technology means that sufficient material can be ob-
tained for downstream microarray/PCR applications. However the massive
reduction in preantral follicle numbers seen in older women has serious im-
plications for the success of techniques such as in vitro maturation.
Supported by: The Wellcome Trust and St Georges Charitable Founda-
tion.
FEMALE REPRODUCTIVE ENDOCRINOLOGY
P-184
EXPRESSION OF CATHEPSIN B AND EPIDERMAL GROWTH
FACTOR IN EUTOPIC AND ECTOPIC ENDOMETRIAL TISSUES
OF PATIENTS WITH ENDOMETRIOSIS. Y.-H. Koo, C.-H. Kim, J.-
S. Kim, S.-H. Kim, H. Chae, B.-M. Kang. Obstetrics and Gynecology, Col-
lege of Medicine, University of Ulsan, Asan Medical Center, Seoul, Korea.
OBJECTIVE: The study was performed to investigate the expression of
cathepsin B (CB) and epidermal growth factor (EGF) in eutopic and ectopic
endometrial tissues of the patients with endometriosis and to clarify if their
expression in endometrial tissue is associated with endometriosis.
DESIGN: Prospective case-control study.
MATERIALS AND METHODS: A total of 34 women with histologically
confirmed endometriosis were recruited for study group. For controls, 14
women undergoing operative treatment for cervical intraepithelial neoplasia
(CIN) or benign gynecologic conditions other than endometriosis were re-
cruited. Eutopic endometrial tissues of both groups and ectopic endometrial
tissue of study group were collected during the operations. We employed real
time RT-PCR to quantify mRNA levels of CB and EGF in these tissues. Then,
we performed western blot analysis to measure the protein levels of CB and
EGF. Mann-Whitney U-test and Kruscal-Wallis test were used for statistical
analysis.
FERTILITY & STERILITY
RESULTS: The expressions of CB mRNA and its protein were signifi-
with endometriosis than in endometrial tissues of controls (P<0.05). In con-
trast to CB, the expressions of EGF mRNA and its protein were significantly
higher in endometrial tissue of controls than in both eutopic and ectopic en-
dometrial tissues of endometriosis patients (P<0.01). There were no signif-
icant differences in the expressions of both CB and EGF between the eutopic
and ectopic endometrial tissues of endometriosis women.
CONCLUSIONS: These data suggest that the higher expression of CB and
the lower expression of EGF in the eutopic and ectopic endometrial tissues
might be associated with the development of endometriosis. In addition,
the changes or the defects of the eutopic endometrium itself might play a
pivotal role in the histogenesis of endometriosis.
Supported by: None.
P-185
SERIAL MEASUREMENT OF SERUM ANTI-MULLERIAN HOR-
MONE IN WOMEN UNDERGOING LAPAROSCOPIC CONSERVA-
TIVE OVARIAN SURGERY. S. H. Han, B. C. Jee, C. S. Suh, K. C. Kim,
W. D. Lee, S. H. Kim. Department of Obstetrics and Gynecology, Seoul Na-
tional University Bundang Hospital, Seong Nam, Gyonggi, Democratic Peo-
ples Republic of Korea; Department of Obstetrics and Gynecology, College
of Medicine, Seoul National University, Seoul, Democratic Peoples Republic
of Korea; Department of Obstetrics and Gynecology, Hamchoon Womens
Clinics, Seoul, Democratic Peoples Republic of Korea; Department of
Obstetrics and Gynecology, Maria Infertility Hospital, Seoul, Democratic
OBJECTIVE: To evaluate the ovarian reserve changes after laparoscopic
conservative ovarian surgery. We prospectively evaluated pre- and postoper-
ative serum levels of anti-mullerian hormone (AMH), and preoperative ovar-
ian volumes using the 3-D ultrasound.
DESIGN: Prospective study.
MATERIALS AND METHODS: Twenty women with a benign ovarian
mass participated (mean  SD: 33.4  7.2 year old, range: 2145 years) after
providing an informed consent. Ovarian masses were removed using a lapa-
roscopic stripping method (ovarian cystectomy). Postoperative pathologies
revealed endometrioma (n 13), mature cystic teratoma (n 6) and mucin-
ous cystadenoma (n 1). Seven patients had bilateral ovarian masses. Serum
values of AMH were measured four times; before operation, one week, one
month, and three months period after operation. Ovary and ovarian mass vol-
umes were measured by 3-D ultrasound before operation. Non-diseased ovar-
ian volumes were estimated by subtracting ovarian mass volumes from
ovarian volumes. AMH data were compared nonparametrically using the
Wilcoxons signed rank test. Correlations between age, non-diseased ovarian
volume, and changes in ovarian reserve were calculated using the Spearman
correlation method.
RESULTS: Mean AMH level was 2.92  1.94 ng/mL before operation but
reduced to 1.18  1.07 ng/mL, at one week postoperatively (P<0.001) and
then slightly increased to 1.61  1.43 ng/mL after one month period
(P0.001) and 1.76  1.91 ng/mL after three months period (P0.016).
However, it subsequently failed to increase to the preoperative level
(P0.023). AMH values were reduced in all 20 patients at 1 week postoper-
atively, and then increased at one to three month postoperatively; moreover,
these increases were found to be positively correlated with total ovarian vol-
ume (r 0.779, P0.002) and diseased ovarian volume (r 0.782,
P0.003) prior to operation.
CONCLUSIONS: Laparoscopic cystectomy of benign ovarian tumors
may be associated with a reduced ovarian reserve in terms of AMH. More-
over, the increased levels of AMH over the 3 months period postoperatively
correlates with the total ovarian volume and diseased ovarian volume in our
study. Therefore, this pilot study suggests that ovarian reserve can be restored
after ovarian surgery in reproductive women. A further study with a longer
follow-up is strongly recommended.
Supported by: None.
P-186
HUMAN CHORIONIC GONADOTROPIN EXHIBITS A DIRECT
DOWNREGULATORY EFFECT ON THE EXPRESSION OF THE
INHIBITORY TYPE II INTERLEUKIN 1 RECEPTOR IN HUMAN
ENDOMETRIAL CELLS. A. Akoum, C. V. Rao, C. Herrmann-Lavoie.
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