530 Autotransplantation of human germ cells

529 IMPACT ON MALE FERTILITY EPIDIDYMITIS MODEL DUE Ziani M., Hakami CHU Amiens,

IN A MOUSE TO CHLAMYDIA

E, Gastaud O., Fourmarier

Urology-Transplantation.

EXPERIMENTAL TRACHOMATIS

ACUTE

530 AUTOTRANSPLANTATION Sofikitis

N., Giannakis

Ioannina

University,

OF HUMAN D., Chatzikyriakidou

GERM

CELLS

A., Baltogiannis

D., Tsambalas

Amiens,

France

INTRODUCTION & OBJECTIVES: The clinical manifestations of Chlamydia Trachomatis (CT) infection in men are urethritis and epididymitis. The development of chronic epididymis and impairment of fertility are possible complications, An experimental model of epididymitis in mice was developed in our laboratory It was shown that CT multiplies within the epididymitis from day 7 after inoculation: the maximum happened on day 11 followed by a decrease until day 30 and a disappearance at day 65. A diffusion of the germ to the testis and the urethritis was noted. Inflammation is present from day 4 to day 11 and decrease from day 15 to day 30 with minimal sequella such as scarring and mononuclear cells infiltrates. The present study was undertaken to evaluate the impact of this experimental epididymitis on male fertility in mouse. MATERIAL & METHODS: a) Infection of mice: C3H male mice 8 weeks old were surgically inoculated in the two epididymis under light anaesthesia. Human genital strain (Bour serovar E) CT was prepared from infected McCouy cells. Mice received in each epididymis 10’ IFU in 40 pL. Control mice were inoculated with the same volume of non infected cells. b) Fertilitv study: Mice were divided into three groups (A, B, C) with in each of them 10 infected mice and 2 controls. Each male was mated with a fertile non infected female and put in a cage 7 days after CT inoculation (group A), 30 days (group B), 90 days (group C). Pregnancies and numbers of young were noted though 3 months. c) Suermoaramm: right and left epididymis of 8 mice (6 infected and 2 controls) were dissected in a 5 cm Nunc dish with Eagle medium and homogenized. A smear was done with 200 pL fixed and Harris-Schorr stained in order to study the spermatozoon morphology. Number and motility studies were done with an in viva examination. RESULTS: The mean number of youngs observed after 3 months infected mice that in the control and there was a decrease of fertility female are put together far away from the out set of infection. The showed a decrease of motility 30 days after inoculation but no modification and no “jerk phenomenon” was noted.

is lower in the when male and spermogramm morphological

CONCLUSIONS: Experimental epididymitis with CT induce a partial infertility a decrease of spermatozoon motility. More yorks are necessary to obtain conclusive information.

with more

Department

of Urology,

loannina,

Greece

INTRODUCTION & OBJECTIVES: We attempted to preserve fertility potential in men with testicular cancer undergoing chemotherapy by transplanting frozen/thawed testicular germ cells post-chemotherapy. MATERIAL & METHODS: Thirty-one men with unilateral testicular malignant disease underwent unilateral radical orchiectomy. At the time of unilateral orchiectomy germ cells were isolated from the contralateral healthy testicle. Biopsies from the contralateral testicle demonstrated absence of neoplasia in all patients. Fractions of testicular germ cells from the healthy testicle were then frozen. Eight out the above 3 I patients received the same standard chemotherapy protocol (for clinical low-volume stage II non-seminomatous germ cell tumours) post-orchiectomy. Six months after the end of the protocol of the administration of the chemotherapeutic drugs (Bleomycin, etoposide, and cisplatin) all eight men were considered to be free of the malignant disease and were found to be azoospermic. At that time the frozen/thawed germ cells were transplanted back to the rete testis of the contralateral to the neoplasia testicle in each of four patients. Rete testis transplantation was ultrasonographically guided. RESULTS: Thirteen months post-transplantation the four men showed sperm concentrations of 1x106/ml, 9 xlOVml, 4x106/ml and 5 x106/ml, respectively, percentage of motile spermatozoa 13%, 2%, 8% and lo%, respectively, and percentage of morphologically normal spermatozoa (WHO criteria) equal to 12%, 22%, 11% and 20%, respectively. In contrast, at the same period after the end of chemotherapy, the four patients who had received chemotherapy but did not undergo autotransplantation were still azoospermic. Twenty months posttransplantation all the four men who had undergone transplantation remained positive for spermatozoa in their ejaculates. In contrast, at the same period after the end of chemotherapy, none of the four men who had received chemotherapy, but did not undergo transplantation, became positive for spermatozoa. CONCLUSIONS: It appears that autotransplantation germ cells post-chemotherapy can result in colonization production of semen samples of sufficient quality technology.

of testicular frozen/thawed of the human testicle and for assisted reproduction

531 GERM CELL DIVISION ANEUPLOIDY SPERMATOGENESIS OF MEN WITH Huann

W., Chen K.K.,

Chang

AT INDIVIDUAL MEIOSIS IDIOPATHIC AZOOSPERMIA

IN

532 TISSUE OUTCOME ASSISTED

Yang-Ming

University,

PERFUSION ESSENTIAL FOR SPERMATOGENESIS OF TESTICULAR SPERM EXTRACTION (TESE) REPRODUCTION

AND FOR

L. Herwin

National

S.

M., Saint F., Petit J.

Department

of Urology,

Taipei,

R.‘, Tosun K.‘, Pinggera

G-M.‘;

Gozzi C.‘, Illmensee

K.*, Bartsch

G.’

Taiwan

INTRODUCTION & OBJECTIVES: An increased aneuploidy has been noted in developing germ cells of men with idiopathic infertility. By using 3-color (chromosome 18, X and Y) fluorescent in situ hybridization (FISH), individual aneuploidy rate in mitotic or meiotic germ cells could be calculated. However, it is not possible to analyze the non-disjunction rate of individual meiotic division based only on FISH signals, because the products from meiosis-I (secondary spermatocyte) or meiosis-II (spermatid) are both haploid cells. The only way to tell them apart are by DNA content or by size of the nuclei. Concurrent using of DAPI-II counter-staining, the nuclear size of the germ cells were also analyzed. Secondary spennatocytes have nuclei larger than 7 mm, while the spermatids are smaller. Therefore, we are able to understand the quality of spermatogenesis focusing on each individual meiotic division by calculating the aneuploidy rate in smaller haploid cells (non-disjunction in meiosis-I) and the aneuploidy rate in larger haploid cells (non-disjunction in meiosis-II), or the ratio of small-to-large cell number (the meiosis-II efficiency). MATERIAL & METHODS: Testis biopsy specimens obtained from 30 azoospermic men were recruited in this analysis. Ten of them are hypospermatogenic (Hype), 20 are maturation arrest (MA). Other tissues from 5 obstructive azoospermic (OA) men were served as control. The FISH efficiency was above 95%. Two independent observers were blinded to the pathological diagnosis. RESULTS: Totally 10,110 cells in 262 tubules were scored. For the haploid cells, the meiosis-II efficiency by small-to-large cell number ratio was 1.32 in OA, 0.3 1 in Hype and 0.34 in MA group. The aneuploidy rates in meiosis-1 (large nuclei) were 27% in OA, 22.4% in Hypo and 22.7% in MA. While the aneuploidy rates in meiosis-II (small nuclei) were 1.8% in OA, 4.7% in Hypo and 5.2% in MA (P
IUniversity Innsbruck,

of Innsbruck, Department Department of Gynaecology,

INTRODUCTION & OBJECTIVES: assisted reproduction (ICSI). In order to perfusion in a single testicle and if the perfusion we collected testicle tissue for perfusion.

of Urology, Innsbruck,

Innsbruck, Austria

Austria,

*University

of

Sperm quality is predictive for the outcome of determine if there are areas of major and minor quality of sperm is correlated with quantity of TESE in accordance to the local testicle tissue

MATERIAL & METHODS: A pre- and intra-operative perfusion mapping was performed using contrast enhanced high resolution Colour Doppler ultrasound. A needle was placed in areas of best and worst perfusion. Perfusion was determined measuring tissue perfusion units (TPU’s) with a Laser Doppler scanner. Tissue was biopsied for TESE and sperm were selected and prepared for ICSI using standard technique. RESULTS: embrionic outstanding.

Best sperm development

CONCLUSIONS: should be replaced Testicular

quality was found m areas of high perfusion following ICSI after perfusion-controlled

(table). The TESE were

To ameliorate the outcome of ICSI, random biopsies by perfusion dependent collection of testicular tissue.

for TESE

biopsy”

Sperm morphology normal abnormal -head** -midpiece -tail” elongated spermatids total sperm evaluated

1 left 1 (70+) 1 64 (72.3%)

1left 2 (lo+) / 2 (13.3%)

right 1 (40+) right 2 (20+) I34 (59.6%) 1 14 (4.0%)

8 (9.1%) 10 (11.4%) 4 (4.5%) 1 2 (2.3%) 88

5 (33.4%) 3 (20.0%) 2 (13.3%) 13 (20.0%) I15

7 (12.3%) 8 (14.0%) 5 (8.8%) 13 (5.3%) I51

* for each testicular sample, 4 microdrops served for sperm evaluation nl), **incl. double and pin, “incl. double and short, +TPU’s European

Urology

Supplements

3 (2004)

6 (17.1%) 7 (20%) 5 (14.3%) 1 3 (8.6%) I 35 (4x25 pl = 100

No. 2, pp.

135