International Journal of Reproductive BioMedicine
Volume:1 Issue: 1, Nov 2003

Spermatids are the earliest male germ cells with one set of haploid chromosomes. Afterexperiments, mainly in rodents, the spermatid injection was introduced in human assistedreproduction to the treatment of men with non-obstructive azoospermia. Spermatid injection is atechnique with particular difficulties that may negatively influence the outcome. The identification,isolation and the assessment of viability, especially for round spermatids, require intensive workand considerable experience. Up to date, it appears that the rates of fertilization and implantationwith round spermatid injection are dramatically low and significantly less compared to the use ofelongated spermatid injection. The extremely low fertilization potency of the round spermatids ledto attempts for their in-vitro culture and maturation. The immaturity of round and elongatedspermatids has raised concerns regarding the potential genetic risk for the offspring. Under thesefacts, a reconsideration of the use of spermatids in assisted human reproduction is necessary.

Spinal cord injury (SCI) has a significant impact on male reproductive functionswhich may lead to infertility. A large number of spinal cord injured men suffer from impairedspermatogenesis. Currently, in vivo gene transfer of molecules with potential therapeutic value hasbeen recognized as a viable method for inducing functional recovery after SCI. This studycharacterized the role of adenovirus-mediated gene transfer into experimentally injured spinal cordsof rats on possible restoration of spermatogenic cell lines.

Young adult Sprague-Dawley rats (200-250g) were assigned into one ofthe three different groups of control, SCI, and adenovirus transfer (Ad) (n=3/ group). Control ratsreceived no injury, nor any surgery. For SCI rats, SCI was produced by a 10g brass rod with a tipdiameter of 2 mm which was dropped from a height of 12.5 mm onto exposed spinal cord at level ofT10 with NYU impactor. Animals were perfused transcardially 43 days post SCI. Both spinal cordand testicular tissues were cryo-sectioned and ultra thin-sectioned, respectively. Cellularmorphology and morphometry were done for spinal cord tissues. The testicular samples wereprocessed for both light and transmission electron microscopy (TEM). The third group of ratsunderwent SCI first, followed by microinjection of LacZ adenoviral vectors (5x106 p.f.u./ μl) alongthe T6-T10 dorsal root entry zone bilaterally. The immune system of animals were suppressedbefore the Ad administration. Each Ad injection was done using a glass micropipet and a Nonojectinjector. Rats were killed 43 days after Ad injections, and the tissues were studied as for othergroups.

The spinal cord lesion extents for SCI and Ad groups were 8.1±3 and 5.8±2.2 mm,respectively (p<0.05). The testicular tissue of controls revealed a normal arrangement ofspermatogenesis cell types. However, impaired spermatogenesis including vacuolization of germcells along with incomplete spermatogenesis were noted in the tubles of SCI group. Also, nucleiand cell membranes of spermatozoa were damaged. In Ad rats, relatively active spermatogenesis,ranging from reappearance of proliferating spermatogonia to the presence of mature spermatozoawere observed in some seminiferous tubles.

Bilateral adenovirus-mediated gene transfer into experimentally injured spinal cords ofrats can restore the ultrastructure of spermatogenesis including mature spermatozoa.

Embryonic stem cells (ESc) are pluripotent cells which have been used as a model tostudy the mechanism that control the embryogenesis and early mammalian development in vitro.The aim of this study was to isolate and produce embryonic stem cells from late blastocyst stageembryos in mice.

Blastocyst stage embryos from pregnant NMRI mice were obtained andcultured for 24 h in DMEM medium. 4-6 days after hatching, the inner cell masses (ICM) formedcolonies which were then collected mechanically and trypsinized. Several subcultures were preparedin the medium supplemented with 0.1 mM 2 Mercaptoethanol, 1000 U/ml Leukemia InhibitoryFactor (LIF) and 10% Fetal Bovine Serum (FBS). The (ESc) were recognized by alkaline phosphateshistochemistry using azo-coupling method.

The results demonstrated that a highly pluripotent stem cell line was derived from theblastocyst stage embryos of NMRI mice; however, the rate of colonies was as low as 10%.

The LIF is effective to culture and maintain the isolated ICM colonies inundifferentiated condition in the absence of feeder layer.

Ovarian functional cyst is one of the most common pelvic mass in reproductive agewhich mostly resolves spontaneously. Sonography is a valuable tool for diagnosis of benign cystwith high accuracy. The objective of this cross sectional study was to evaluate the accuracy oftransvaginal sonography in detecting type of ovarian cyst and compare the results wieh cytologicalresults.

82 women in reproductive age who have had simple ovarian cysts withbenign criteria which unresolved after taking contraceptive pills for 3 months were considered forthis clinical study. Transvaginal ultrasound-guided aspiration of cysts were done and were then sentto the pathological evaluation. Also, all data regarding the size of the cysts and aspirated fliud wererecorded in charts for further statistical analysis.

The accuracy of transvaginal ultrasound comparing with cytology on diagnosis forfunctional cysts was 94.9%, for epithelial ovarian cyst was 97.5% and for endometrioma was 97.5%(P= 0.0001).The size of cysts with diameter of <10cm was not related to the quality of cysts.

The results showed that sonography is a valuable and reliable tool for diagnosis ofbenign ovarian cyst. It seems that if a mass appears benign by ultrasound morphologic criteria,probability of it being malignant is near to zero, which can be aspirated by transvaginal routewithout any fear from missing of malignancy or complication.

The aim of this study was to determine the correlation between ultrastructural studiesfor pinopodes expression after ovarian hyperstimulation and progesterone injection in mice.

Adult NMRI mice were superovulated using human menopasualgonadotropic (hMG) and human chorionic gonadotropic (hCG) hormones; after that, daily injectionof progesterone (1 mg/mouse) was performed. Animals were sacrificed by cervical dislocation 3.5and 4.5 days after hCG injection. Tissues of uterine horns were obtained and processed for scanning(SEM) and transmission (TEM) electron microscopy studies. The pseudopregnant control sampleswere studied same as experimental groups.

The SEM and TEM observations showed that in control groups on 3.5 days of pregnancy,there were some pinopodes. All apical cell surfaces expressed these projections on the forth day. Inprogestrone-injected group, well developed pinopods were expressed 3.5 days after hCG injectionand they were transformed to small projections on the fourth day following hCG injection. Also, thelife span of pinopods was limited to a short time. At the TEM levels, the pinopods were seen asswelling process on the apical surface, which were more pronounced on day 3.5 of hCG injection inhyperstimulated and progestrone injection.

The progestrone may cause premature expression of pinopodes and the implantationfailure after ovarian induction may be due to these timing changes.

While traditional semen parameters are of significant clinical value, total fertilizationfailure in IVF cycles is not uncommon. Sperm function testing such as; Hamster egg penetrationtest has severed limitations as a clinical test. The aim of this study was to evaluate the predictivevalue of semen parameters by using of intracellular calcium [Ca2+]I increase in response toprogesterone.

The [Ca2+]i response to progesterone was measured in spermatozoa of86 patients referring to the Assisted Conception Unit for semen analysis. The patients were dividedinto 3 groups; according to their semen parameters and measured intracellular [Ca2+]i increasing inresponse to progesterone.

There was no significant correlation between each individual semen parameter and[Ca2+]i elevation in response to the progesterone, but most of the patients in each group had[Ca2+]i increasing as expected based on sperm parameters. However, there were cases in groups 1and 2 (Normal and IVF) that demonstrated [Ca2+]i increases which were poor or lower thanexpected. Out of the 22 patients in the normal category, 8 cases had poor response to [Ca2+]Iincrease and out of the 47 patients in the IVF group, 9 patients were as well. In addition wemeasured [Ca2+]I increases in 6 fertile donor samples for comparison purposes.

[Ca2+]i increase in response to progesterone is related to predicting value of spermparameters in most cases. However, the response of sperm to progesterone could be different insome cases that are expected in normal or IVF category based on our semen analysis criteria. Wesuggest that the [Ca2+]i measurements may perfect the sperm fertility potential.

Intrauterine insemination (IUI) is generally attempted before proceeding to moreexpensive and invasive assisted reproductive techniques such as invitro fertilization (IVF) andintracytoplasmic sperm injection (ICSI). This procedure is most commonly performed as atherapeutic method for couples with a wide variety of subfertility etiologies, such as low count orlow motility of sperm, or an incompatibility between the sperm and the cervical mucus. Theobjective of this clinical trial study was to compare the correlation between the semen parametersand pregnancy rates in patients undergoing hyperstimulation and IUI.

336 infertile couples that underwent 336 cycles of IUI with washedhusband’s semen were included in this study. All patients’ charts were reviewed for age, etiologyand duration of infertility, semen characteristics and pregnancy rates. The SPSS 9 software and Chisquaretests were applied for statistical analysis. P<0.05 was determined as statistical significance.

Total pregnancy rates were18.2% (61 out of 336 cycles). Postwash semen parametersincluding: sperm count ≥10× 106, motility ≥50% (grade III and IV >20%) had significant effect onpregnancy rates after IUI. The Outcome of this procedure was not significantly affected by femaleage, duration or etiology of infertility.

Postwash semen quality was the most important factor for predication of successfulpregnancy in this study.

The high fertilization failure after IVF treatment cycles could be related tochromosomal abnormalities. This study was carried out to assess the frequency of chromosomalabnormality on human oocytes lacking signs of fertilization 18-20 h after insemination.

On day one, 18-20 h after insemination (IVF), fertilization wasconfirmed when two pronuclei (normal IVF) or more pronuclei (poly pronucleus FR) were present.Chromosomal analysis of unfertilized oocytes was carried out within 20-24 h of collection. Alloocyte did not sign of pronuclei were collected from total fertilization failure, TFF (FR=0) or partialfertilization failure, PFF (FR=10-90%). Chromosomal preparation was carried out as described byTarkowski’s techniques. The average of finding between two groups was compared by X2 test.

Chromosome spreading permitted adequate analyzing in 348 unfertilized oocytes. In33.6% chromosomal aneulpoidy was observed with the following frequencies; hypo-hyploidy,22/348 (6.4%), hyper-hyploidy, 42/348 (12.2%) and diploidy, 52/348 (15%). The frequency ofaneuoplidy was significantly higher in TFF group 33/80 (41%) than PFF group 83/268 (31%),p<0.01, X2. The most frequent numerical aberration was observed in chromosome group, G of thehuman karyotyped.

Since cytogentic analysis of failed fertilized oocytes and sperm function tests are veryhelpful for direct information on low success rate of fertilization, further studies analyzing on bothgametes function in TFF cycles will be needed.