used for comparison purposes among the various specimens evaluated. TheTFSF (x106) was calculated as count (x106) x % motility x % normalmorphology.

Results: The average normal morphology of normozoospermic specimensbefore processing was 41.0�8.1, which was significantly different formteratozoospermic specimens (P�0.05). The average normal morphology forunprocessed teratozoospermic specimens classified as mild, moderate andsevere was 24.0�3.0, 15.2�3.0 and 6.4�2.0, respectively. Significantdifferences were noted between each classification (P�0.05). The recoveryof the TFSF for normozoospermic specimens was 24.0�18.8. The recoveryof TFSF for mild, moderate and severe teratozoospermia was 17.2�1.4,9.4�6.6 and 2.9�2.4, respectively. Significant differences were noted be-tween each classification (higher than 100x106), a definite trend was notedfor the sperm count to decrease when the severity of the morphologicalabnormalities increased.

Conclusion: Sperm morphology in mild and moderate teratozoospermiacan reach an acceptable level for normality after processing. On average,sufficient numbers of morphologically normal spermatozoa can be har-vested to perform conventional IVF insemination. However, although thosenumbers were high enough to obtain optimal fertilization rate as observedin normozoospermic specimens, the actual improvements in fertilizationrates and embryo development must be established. Our results provideencouraging data to optimize the potential improvement of the teratozo-ospermic factor and the use of the TFSF to standardize the morphologicalfactor in the insemination doses, which are usually calculated based on themotile sperm fraction.

P-270

Effect of the quality and origin of sperm on fertilization, zygote mor-phology, early cleavage and pregnancy outcome after ICSI. Eun JeongOh, Il Kyung Jeon, Chung Won Kim, Kwang Rae Kim, Sung Il Roh, HyunSoo Yoon. Infertility research Ctr, MizMedi Hosp, Seoul, Republic ofKorea.

Objective: To investigate the effect of the quality and origin of sperm onzygote morphology, first cleavage time and pregnancy outcome after ICSI.

Design: Study groups were divided into five according to the quality andorigin of sperm. A; normal (control, n�415 embryos/ 97 cycles), B; mildoligoasthenoteratozoospermia (OATs, n�536 embryos/ 127 cycles), C;severe OATs (n�149 embryos/ 38 cycles) in the ejaculated sperm groupand D; testicular sperm from obstructive azoospermia (n�45 embryos/ 11cycles), and E; testicular sperm from non-obstructive azoospermia (n�38embryos/ 8 cycles).

Materials and Methods: A total of 341 cycles undergone ICSI from May2002 to March 2003 at MizMedi Hospital were studied. Zygote morpholo-gy(size difference, average size, nucleoli patterns of PN and cytoplasmichalo) were compared. Nucleoli patterns were subdivided into three groups;symmetric, asymmetric, and abnormal pattern. The first cleavage time andembryo development were evaluated at 24 to 26 hours and on day 3 afterICSI, retrospectively. Clinical pregnancy and implantation rate were inves-tigated.

Results: Patient age and zygote morphology except nucleoli patterns ofPN did not differ among the groups. Fertilization rates were higher ejacu-lated sperm groups than testicular sperm groups. Zygotes with a symmetricnucleoli or early cleavage showed a significantly higher developmental ratethan the others. The ratio of zygotes with a symmetric nucleoli pattern orearly cleavage embryos did not differ among the groups except group E. Theratio of good embryos arose from zygotes with a symmetric nucleoli patternor early cleavage and pregnancy outcome did not differ among the groupsexcept group E. However, in group E, the ratio of good embryos wassignificantly lower than in the other groups although the zygotes showed asymmetric nucleoli pattern or early cleavage. Pregnancy outcomes of groupE were also poorer than the other groups; however, there was no statisticallysignificant difference.

Conclusion: In this study, the quality and origin of sperm, especiallytesticular sperm from non-obstructive azoospermia, affected not only nu-cleoli pattern of zygote morphology and first cleavage time but also furtherembryo development and pregnancy outcome, although the zygotes andembryos showed good morphology. In light of this study, we suggest thatdevelopmental competency after fertilization may be affected by somefactors derived from sperm, in the case of non-obstructive azoospermia.

P-271

Treatment of freeze-thawed testicular sperms with Human FollicularFluid (HFF) improves their motility and is useful to select viable spermsfor ICSI procedure. Paris Keynezhad, Maryam Jenabi, Saied Sahe-bkashaf, Vida Feizy, Hamid Sahebkashaf. Navid’s Institute of Infertility,Tehran, Iran (Islamic Republic of).

Objective: The motiltiy of sperms obtained from testicular biopsy isgenerally very low.Since in some cases, it is nessesary to freeze the biopsiedtissue in order to prevent of repeated testis biopsy, and for future use oftesticular sperms in ICSI ,on the other hand, many of these cells may losetheir viability during cryopreservation, it is reasonable to find a way whichmay help us to select viable sperms after thawing, for injecting them intooocytes.

According to previous studies on the positive effects of hFF on spermmotility and its capacitation,we tried to take advantage of this property andto evaluate the motility of freeze-thawed testicular sperms, following cul-ture in different combinations of hFF and Ham’s F-10.

Design: Comparison of testicular sperm motility immediately after thaw-ing ,and following culture in different concentrations of hFF.

Materials and Methods: The biopsied testicular tissue (n�8) was sub-jected to mechanical mincing by two glass slides and recovered testicularsperms were freezed with conventional sperm freeze solution (glycerol15%). After thawing, samples were allocated to four groups as follow:Group 1(Control);Ham’s F-10/without culture,Group 2; Ham’s F-10�0%hFF/with 24 hrs culture, Group 3; Ham’s F-10 �25% hFF/with 24hrsculture, Group 3; Ham’s F-10 �50% hFF/with 24hrs culture.Culture wasdone under 5% CO2 and 37° C .In group 1 (Control), motility was evaluatedimmediately after thawing .In Group 2,evaluation was done to determine ifculture alone has a positive effect on motility and in remainig groups(3 and4), the effect of different concentrations of hFF on testicular sperm motilitywas determined. At least 100 sperms were counted for motiltiy scoring ineach group.

Statistical analysis was performed by paired t-test between control andexperimental groups ,and a comparison of Mean�SD for motility betweengroups 2,3 and 4 was done to define the best concentration of hFF fortesticular sperm culture after thawing. P�0.05 considered significant.

Results: Sperm motility immediately after thawing was 3.00�4.40%(Group 1).After 24 hrs of culture, motility was 21.25�12.61%(Ggroup2),30.75�16.23% (Group 3) and 43.0�17.00% (Group 4) respec-tively.The differences between control and other groups(cultured in FF) wasstatistically significant. Also the motility was significantly higher in group4 compared with other groups(2,3).

Conclusion: These results suggest that 24 hrs culture of freeze-thawedtesticular sperms in media containing 50% hFF may improve its motility,and, in addition of helping us to select viable sperms for ICSI, may bebeneficial for these cells in aspect of optimizing their quality and ability tofertilize oocytes.

S212 Abstracts Vol. 80, Suppl. 3, September 2003