BACKGROUND:Cyclophosphamide (Cy), a widely used anticancer drug, is associated with significant testicular damage and sterility. Co-administration of the immunomodulating compound AS101 during chemotherapy treatments was previously shown to protect organs against cytotoxic damage, without attenuating the drug's anticancer effect. In this animal study, we investigated the effect of AS101 on testicular damage, sperm DNA damage and infertility induced by Cy. Akt and glycogen synthase kinase-3beta (GSK-3beta) phosphorylation were investigated as a possible chemoprotective mechanism.METHODS:Mature male mice, 10 in each group, were injected intraperitoneally with 200 mg/kg Cy once a week for 5 weeks, with or without concurrent treatment with 10 microg per mouse AS101 three times per week. Damage to testicular tubules and sperm production was determined, sperm chromatin damage was analyzed and fertility was gauged. Akt and GSK-3beta phosphorylation were evaluated.RESULTS:Co-treatment with AS101 during the course of Cy administration significantly reduced the percentage of damaged seminiferous tubules (76.0 +/- 10.8% versus 40.3 +/- 2.6%), and reduced sperm DNA fragmentation (%DFI) from 44.7 +/- 1.0% to 25 +/- 6.5%. Co-treatment with AS101 also partially protected against the decrease in numbers of impregnated females and litter size. AS101 increased Akt and GSK-3beta phosphorylation.CONCLUSIONS:Our results indicate that AS101 can significantly protect against Cy-induced testicular damage and sperm DNA damage, probably by acting through Akt/GSK-3beta phosphorylation.

Poor spermatogenesis in patients with inflammation of the genital tract is associated with scrotal hyperthermia. These patients can benefit from acupuncture treatment. We conducted a study to verify whether the influence of acupuncture treatment on sperm output in patients with low sperm density is associated with a decrease in scrotal temperature. The experimental group included 39 men who were referred for acupuncture owing to low sperm output. The control group, which comprised 18 normal fertile men, was used to define a threshold (30.5 degrees C) above which scrotal skin temperature was considered to be high. Accordingly, 34 of the 39 participants in the experimental group initially had high scrotal skin temperature; the other five had normal values. Scrotal skin temperature and sperm concentration were measured before and after acupuncture treatment. The five patients with initially normal scrotal temperatures were not affected by the acupuncture treatment. Following treatment, 17 of the 34 patients with hyperthermia, all of whom had genital tract inflammation, had normal scrotal skin temperature; in 15 of these 17 patients, sperm count was increased. In the remaining 17 men with scrotal hyperthermia, neither scrotal skin temperature nor sperm concentration was affected by the treatment. About 90% of the latter patients suffered from high gonadotropins or mixed etiological factors. Low sperm count in patients with inflammation of the genital tract seems to be associated with scrotal hyperthermia, and, consequently, acupuncture treatment is recommended for these men.

OBJECTIVE: Up to 10% of couples who visit a fertility clinic for treatment receive a diagnosis of unexplained fertility (UI) due to absence of a correctable abnormality. Using Sperm Organellar Morphology Examination (MSOME), we tried for the first time to verify whether males with UI may exhibit nuclear impairments in their spermatozoa (nucleo-teratozoospermia, NT), undetected by the routine LM. In addition, we aimed to evaluate the benefit of Intracytoplasmic Morphologically-selected Sperm Injection (IMSI) for couples with UI and NT. DESIGN: At least 7% of spermatozoa with normal nucleus per ejaculate is the laboratory standard for normal fertile males. This threshold enabled comparison between couples with UI, who were and were not defined as NT. MATERIALS AND METHODS: Ninety UI couples with a normal andrological profile and a normal female partner (≤38 years), who failed to conceive after four COH/IUI trials and at least 1 standard IVF and intracytoplasmic sperm injection (ICSI) in sibling oocytes with fertilization rate ≥ 50% in IVF. Each of these couples was referred for further evaluation by MSOME and a single IVF-IMSI trial, were sperm selection is based on MSOME. Unvaried analysis of variance (ANOVA) for continuous variables and Chi square tests for discrete variables were used. RESULTS: Sixty five (72.2%) of the 90 males with UI were defined by MSOME as NT. The males with and without NT (n=65 and n=25, respectively, were statistically similar in their demographic data and morphological normalcy of the sperm cell subcellular organelles, except percentage of normal nucleus, which was significantly lower in males with NT compared to those without NT (2.6±2.3% vs 15.4±6.2%, F=208.1, z=−7.3, p≤0.01). Comparison between the two study groups in IVF-IMSI outcome parameters revealed no statistical difference between them in any of these variables except implantation and pregnancy rates, which were significantly higher in the NT group compared to those without NT (25.1±37.0% vs. 5.3±15.0%; F=6.7; and 64.6% vs.12.0%; Chi square=20.0, respectively, p≤0.01). CONCLUSIONS: MSOME is a first rate tool for revealing a hidden male fertility factor of infertility in couples with unexplained infertility. IVF–IMSI should be considered for UI couples who have been diagnosed by MSOME to have nucleo-teratozoospermia.

Campylobacter fetus subsp. fetus infection can occur in female sheep, causing infertility or abortion. Despite extensive research on the effect of these bacteria on female fertility, little research has been done on the influence of C. fetus subsp. fetus on the male factor. Our objective was to examine the influence of C. fetus subsp. fetus on ram sperm. Motility index, percentage of live spermatozoa, mean alphat value (an indication of the chromatin stability of the sperm cell) and percentage of sperm cells expressing the FAS receptor were measured in sperm incubated in the presence or absence of C. fetus subsp. fetus. The motility index and viability of sperm incubated with the bacteria were lower than those of untreated sperm samples after 5 h. In bacteria-incubated sperm cells, the percentage expressing FAS receptor was already significantly elevated at 15 min. Bacteria-incubated sperm showed a greater prevalence of morphological damage. The bacteria were attached to tail and acrosome regions, and the sperm damage was concentrated in both the motility and chromatin regions. Bacteria-infected sperm cells showed a decrease in motility, increase in early acrosome reaction and chromatin damage. Similar effects were induced by incubation of the sperm with supernatants from C. fetus subsp. fetus cultures. Thus this study demonstrates that C. fetus subsp. fetus has a detrimental effect on the quality of ram sperm.

OBJECTIVE:Incubation of ejaculated spermatozoa at 37 degrees C is recommended for IVF-intracytoplasmic sperm injection. Preselection of sperm cells with morphologically normal nuclei before microinjection, adapted in our laboratory, is usually a time-consuming procedure. Therefore, we aimed to verify whether incubation at 37 degrees C could affect the morphologic integrity of sperm nuclei. DESIGN:Time-kinetics studies testing fine morphology of the sperm nuclei upon prolonged in vitro incubation. SETTING:Male Fertility Laboratory at Bar-Ilan University, Ramat Gan, Israel. PATIENT(S):Forty-two males selected at random, who were referred for sperm preselection before ICSI. MAIN OUTCOME MEASURE(S):Morphologic integrity of the sperm nuclei, obtained by motile sperm organelle morphology examination. RESULT(S):After 2 hours of incubation at 37 degrees C a significant decrease occurred in the morphologic integrity of the sperm nuclei, compared with the initial state (4.7% +/- 2.8% vs. 6.8% +/- 3.5%, t = 3.2, Por=2 hours) sperm manipulations for assisted reproduction therapy should be performed at 21 degrees C rather than 37 degrees C.

This current study investigated a link between stress and coping with some male infertility problems via psychoneuroimmunological theories. The study was carried out on 77 males who came to a fertility clinic for sperm diagnosis. Appraisal and coping with stress reaction to negative life events and to infertility were related to non-sperm cells within the seminal plasma. Those men who had felt strained from external events and from problems of infertility, and had used emotional escape coping strategies, were more prone to have an immunosuppressed reaction, indicated by bacteria within their plasma. Those using task focused control strategies were more prone to have an immunostimulatory reaction, indicated by white blood cells, including in some cases, sperm destroying phagocytes suggesting a possible auto-immunity reaction. When the analyses were done separately on patients with known male fertility problems in comparison with those with unexplained infertility, the two opposing psychoneuroimmunological processes became more pronounced.

BACKGROUND:To verify whether or not microinjection of sperm with a normal nuclear shape but large vacuoles affects IVF-ICSI pregnancy outcome.METHODS:A comparative study testing IVF outcome parameters of IVF-ICSI, based on morphological selection of spermatozoa with normal nuclei against those based on microinjection of sperm with a normal nuclear shape but large vacuoles. An experimental group, including 28 IVF-ICSI cycles, where only embryos obtained from microinjection of spermatozoa with a normal nuclear shape but large vacuoles were transferred, was matched with a control group, including 28 IVF-ICSI cycles, where only embryos obtained from microinjection of spermatozoa with a strictly defined morphologically normal nuclear shape and content were transferred. The main outcome was IVF-ICSI pregnancy rate.RESULTS:The experimental group exhibited a significantly lower pregnancy rate per cycle and significantly higher abortion rate per pregnancy compared to the control group (18 versus 50%, and 80 versus 7%, respectively, P=0.01).CONCLUSION:Microinjection of vacuolated sperm appears to reduce the pregnancy rate and appears to be associated with early abortion.

In previous studies, a new IVF method of intracytoplasmic morphologically selected sperm injection (IMSI) was introduced, based on motile sperm organellar morphology examination (MSOME). It was concluded that microinjection of morphologically selected sperm cells with strictly normal nucleus, defined by MSOME, improves IVF-ICSI outcome. The aim of the present study was to confirm this conclusion in new, enlarged study groups. Comparison between 80 couples, who underwent an IVF-IMSI trial, with matched couples, who underwent a standard IVF-ICSI procedure, confirmed that pregnancy rate following IVF-IMSI was significantly higher, and abortion rate significantly lower than in the routine IVF-ICSI (60.0 versus 25.0%, and 14 versus 40% respectively, P :5 0.05). Another comparison was performed between matched IMSI groups, where embryos were obtained from microinjection by spermatozoa with a morphologically normal nucleus ('best' group, n = 70) and a 'second best' group was selected, where embryos were obtained from microinjection of spermatozoa with minimal morphological impairment, since no other sperm cells were available. It was confirmed that microinjection by 'second best 'spermatozoa result in significantly lower pregnancy and delivery rates and significantly higher abortion rates than microinjection with 'best' spermatozoa (25.7 versus 58.2%, P <= 0.01; 17.1 versus 52.8%, P <= 0.01, and 33.3 versus 9.7%, P <= 0.02 respectively). The present study has strengthened previous conclusions.

BACKGROUND:Our preceding studies have already demonstrated the advantage of intracytoplasmic morphologically selected sperm injection (IMSI) over the conventional IVF-ICSI procedure in terms of pregnancy rate. This study was undertaken to determine whether the increased pregnancy outcome was attributable to the preferred nuclear morphology of the selected spermatozoa, and not to the special sperm preparation technique modified by IMSI.METHODS:Comparison between two matched IMSI groups, i.e. negative comprising 38 cycles, where no spermatozoa with intact nuclei were available for microinjection; and positive, involving ovum microinjection by spermatozoa with strictly defined morphologically normal nuclei.RESULTS:Implantation and pregnancy rates were significantly higher, and abortion rates significantly lower, in the positive group compared with the negative group (25.0+/-25.9 versus 5.9+/-12.9%, F=15.8, P< or =0.01; 52.6 versus 18.4%, chi2=9.7, P< or =0.01; and 10.0 versus 57.1%, chi2=7.1, P< or =0.02, respectively).CONCLUSIONS:Implantation and pregnancy by ICSI is associated with morphological nuclear normalcy of sperm. Sperm with a morphologically abnormal nucleus usually have low fertility potential, but some with certain nuclear abnormalities may still be able to produce pregnancy following ICSI.

In-vitro fertilization (IVF) is associated with a significant risk of ovarian hyperstimulation syndrome (OHSS), especially in young women with polycystic ovaries (PCO). One of the strategies to prevent OHSS is 'coasting', i.e: withholding human chorionic gonadotropin (hCG) administration until estradiol drops to a safe concentration. This action could lead to immature oocyte retrieval. Over the last decade, a significant progress has been made in recovering immature oocytes followed by in-vitro maturation (IVM) and in-vitro fertilization. Recently it has been reported that microinjection of morphologically selected spermatozoa, with normal nuclei, into retrieved oocytes, improves significantly the incidence of pregnancy in couples with previous ICSI failure, since sperm morphology could place a crucial role in ICSI outcome. Presentation of a case report of pregnancy, achieved following microinjection of morphologically selected spermatozoa into in-vitro matured oocytes. A 28 years old PCO patient, with 7 previous IVF/ICSI cycles failures, was treated with gonadotrpin releasing hormone agonist (Triptoreline acetate CR, Ferring GmbH, Germany) and a combination of hMG HP (Menopur, Ferring GmbH, Germany) and recombinant FSH (Gonal F, Serono, Switzerland). Since many middle size follicles developed and serum estradiol level was 2045 pg/ml., 'coasting' was performed for two days to prevent OHSS. Five oocytes were aspirated 34 hours post hCG administration, denuded 5 hours post ovum pick up and assessed for meiotic maturity under inverted microscope. Since all oocytes were immature (3 GV and 2 M I oocytes), they were cultured over night in P-1 medium (Irvine Scientific, Santa Ana, CA., USA), at 370c in 5% CO2 and humidified air. Morphologically selected spermatozoa were also incubated over night in the same conditions. 27 hours post ovum pick up, 4 oocytes reached to M II phase and microinjected with motile selected spermatozoa. Embryo transfer was performed 44 hours post ICSI. Three oocytes were fertilized and cleaved (75%). Three embryos were transferred (two of 3 cell and one of 4 cell embryos, grade III). A twin pregnancy developed. Immature oocytes had the ability to extrude first polar body and reach M II stage, even after incubation with ordinary culture medium such as P-1 medium, without supplementation of essential growth factors and hormones. To the best of our knowledge, this is the first report indicating that in-vitro matured oocytes can be fertilized and cleaved with morphologically selected spermatozoa, incubated overnight in the same conditions, and finally achieving a pregnancy.

The objective of this research was to evaluate the influence of the immunomodulator ammonium trichloro (dioxoethylene-0,0') tellurate (AS101) on caspase activity, mitochondrial transmembrane potential (δ Ψm), and DNA stability in human sperm. Cells with Intact δ Ψm, active caspase-3 (CP3), active caspase-9 (CP9), and DNA stability, were measured in washed spermatozoa obtained from nine patients referred to the fertility laboratory after 60 minutes incubation with or without two AS101 concentrations. Sperm samples underwent routine semen analysis (WHO 1992), then washed with Ferticult medium and diluted to a concentration of 2X106 cells/ml. Aliquots were then incubated with either 0.5 μg/ml or 1 μg/ml AS101. A lipophilic cationic dye JC1 (5,5,6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine chloride) was used to detect disrupted or intact δΨm in living spermatozoa. Activated CP3 and CP9 were detected in spermatozoa using the CaspaTag kit (CaspaTag, Intergen Co., Biotest, Israel) according to the manufacturer's instructions. The percentage of active CP positive cells and mean fluorescence was calculated. The sperm chromatin structure assay was conducted according to the method of Evenson and Jost (1994). SCSA parameters used were mean channel of αt distribution and percentage of cells outside the main αt peak (%Compαt). p<0.05 was significant by student's test. The results from the various methods are summarized in the table. Treatment of sperm cells with AS101 significantly reduced CP3 and CP9 activity in the caspase positive population (P<0.05). No significant difference was found between the two concentrations. Treatment of sperm with AS101 had also a significant positive effect on sperm chromatin stability as shown by αt and %Compαt values (p<0.05). however, no significant difference was found in the percentage of cells with intact δΨm and percent of cells exhibiting active cp3 and cp9. Tabled 1 We found that in vitro incubation of sperm with AS101 did not affect the percentage of cells exhibiting apoptotic markers. However, AS101 reduced the activity of CP3 and CP9 in the apoptotic cells and improved DNA stability. This data suggests that the sperm population undergoing an apoptotic process cannot be rescued in vitro. However, the apoptotic process can be slowed down significantly by treatment with AS101. Further research should be done to examine whether AS101 also has other protective effects.

ObjectiveTo examine mitochondrial activity, caspase level and activity, and nuclear chromatin stability in sperm cells of individual males in response to cryoprotectors and the cryopreservation process.DesignNine subjects referred to our laboratory for routine checkups[according to the World Health Organization (1992)] participated in the study. Each fresh semen sample was divided into three aliquots: 1. control (unprocessed); 2. fresh semen with citrate - egg yolk - glycerol (TYB); 3. fresh semen with sperm-freezing medium (SFM). Diluted samples were equilibrated 30 min and cooled in a programmed cooling freezer (Planer Products, Ltd., Perkasie, PA) using two rate protocols, then plunged into liquid nitrogen for two weeks.Materials and methodsThe samples were tested in 3 categories: fresh semen, equilibrium with cryoprotectors, and post-thawing. A lipophilic cationic dye JC1 (5,5,6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine chloride) was used to detect mitochondrial ΔΨm level in spermatozoa. Caspase-3 (CP3) and caspase-9 (CP9) activities were tested using the CaspaTag kit. The average percentage of cells with active CP positive and mean fluorescence was calculated. The sperm chromatin structure assay (SCSA) was conducted according to the method of Evenson and Jost. SCSA parameters used were mean channel of αt distribution and percentage of cells outside the main αt peak (%Compαt).ResultsMitochondria activity of post-thaw spermatozoa decreased in both cryoprotectors: in TYB group to 27% vs. 64 % in control (p<0.05) and in SFM to 32% vs. 64% in control (p<0.05). The average CP3 level increased at the post-equilibrium stage as compared with control: 33% in TYB group, and 38% in SFM. Similarly, the percentage of CP9 increased: 34% in TYB groups and 26% in SFM. The process of post-thawing increased the percentage of cells with positive intensity of CP3 in TYB group by 75%, in SFM by 73%, and that of CP9 significantly increased in the TYB group by 56% and in SFM by 76%. At the post-equilibrium stage, the following level of active CP3 was detected: in the TYB group 1168± 394.5 vs. 811.3 ± 271.1 in control (p<0.01); and in the SFM group 11314.6 ± 389.4 vs. 811.3 ± 271.1 (p<0.01). The level of active CP9 did not change. However, post-thawing, CP3 activity in positive cells significantly decreased to 580.2 ± 207.6 in TYB and to 727.1 ± 223.4 in SFM (p<0.05). CP9 activity significantly decreased in comparison with control (617.2 ± 135.0) to 344.2 ± 59.0 in TYB group (p<0.05) and 471.7 ± 41.6 in SFM (p<0.05). The assay of SCSA shows that the mean αt value in fresh samples was 180.9 ± 14.3, which significantly decreased in all stages of cryoprocesses to 165.8 ± 20.7 (p<0.05) in TYB and 164.7 ± 16.4 in SFM (p<0.05). The percentage of COMPαt cells in the post-thaw did not differ from the control.ConclusionHuman sperm cells exposed to cryoprotectors increase their apoptotic markers both in quantity and in intensity, however, post-thawing, there was further increase in the percentage of cells exhibiting these markers but their intensity levels decreased. The underlining mechanism of this phenomenon in now investigated. Cryopreservation processes have a tendency to increase the quantity of sperm cells expressing apoptotic markers. On the other hand, at stages of equilibrium and post-thawing, increase in intensity of caspases and chromatin stability was found. We will examine the differences between the cryoprotective media by virtue of this study. ObjectiveTo examine mitochondrial activity, caspase level and activity, and nuclear chromatin stability in sperm cells of individual males in response to cryoprotectors and the cryopreservation process. To examine mitochondrial activity, caspase level and activity, and nuclear chromatin stability in sperm cells of individual males in response to cryoprotectors and the cryopreservation process. DesignNine subjects referred to our laboratory for routine checkups[according to the World Health Organization (1992)] participated in the study. Each fresh semen sample was divided into three aliquots: 1. control (unprocessed); 2. fresh semen with citrate - egg yolk - glycerol (TYB); 3. fresh semen with sperm-freezing medium (SFM). Diluted samples were equilibrated 30 min and cooled in a programmed cooling freezer (Planer Products, Ltd., Perkasie, PA) using two rate protocols, then plunged into liquid nitrogen for two weeks. Nine subjects referred to our laboratory for routine checkups[according to the World Health Organization (1992)] participated in the study. Each fresh semen sample was divided into three aliquots: 1. control (unprocessed); 2. fresh semen with citrate - egg yolk - glycerol (TYB); 3. fresh semen with sperm-freezing medium (SFM). Diluted samples were equilibrated 30 min and cooled in a programmed cooling freezer (Planer Products, Ltd., Perkasie, PA) using two rate protocols, then plunged into liquid nitrogen for two weeks. Materials and methodsThe samples were tested in 3 categories: fresh semen, equilibrium with cryoprotectors, and post-thawing. A lipophilic cationic dye JC1 (5,5,6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine chloride) was used to detect mitochondrial ΔΨm level in spermatozoa. Caspase-3 (CP3) and caspase-9 (CP9) activities were tested using the CaspaTag kit. The average percentage of cells with active CP positive and mean fluorescence was calculated. The sperm chromatin structure assay (SCSA) was conducted according to the method of Evenson and Jost. SCSA parameters used were mean channel of αt distribution and percentage of cells outside the main αt peak (%Compαt). The samples were tested in 3 categories: fresh semen, equilibrium with cryoprotectors, and post-thawing. A lipophilic cationic dye JC1 (5,5,6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine chloride) was used to detect mitochondrial ΔΨm level in spermatozoa. Caspase-3 (CP3) and caspase-9 (CP9) activities were tested using the CaspaTag kit. The average percentage of cells with active CP positive and mean fluorescence was calculated. The sperm chromatin structure assay (SCSA) was conducted according to the method of Evenson and Jost. SCSA parameters used were mean channel of αt distribution and percentage of cells outside the main αt peak (%Compαt). ResultsMitochondria activity of post-thaw spermatozoa decreased in both cryoprotectors: in TYB group to 27% vs. 64 % in control (p<0.05) and in SFM to 32% vs. 64% in control (p<0.05). The average CP3 level increased at the post-equilibrium stage as compared with control: 33% in TYB group, and 38% in SFM. Similarly, the percentage of CP9 increased: 34% in TYB groups and 26% in SFM. The process of post-thawing increased the percentage of cells with positive intensity of CP3 in TYB group by 75%, in SFM by 73%, and that of CP9 significantly increased in the TYB group by 56% and in SFM by 76%. At the post-equilibrium stage, the following level of active CP3 was detected: in the TYB group 1168± 394.5 vs. 811.3 ± 271.1 in control (p<0.01); and in the SFM group 11314.6 ± 389.4 vs. 811.3 ± 271.1 (p<0.01). The level of active CP9 did not change. However, post-thawing, CP3 activity in positive cells significantly decreased to 580.2 ± 207.6 in TYB and to 727.1 ± 223.4 in SFM (p<0.05). CP9 activity significantly decreased in comparison with control (617.2 ± 135.0) to 344.2 ± 59.0 in TYB group (p<0.05) and 471.7 ± 41.6 in SFM (p<0.05). The assay of SCSA shows that the mean αt value in fresh samples was 180.9 ± 14.3, which significantly decreased in all stages of cryoprocesses to 165.8 ± 20.7 (p<0.05) in TYB and 164.7 ± 16.4 in SFM (p<0.05). The percentage of COMPαt cells in the post-thaw did not differ from the control. Mitochondria activity of post-thaw spermatozoa decreased in both cryoprotectors: in TYB group to 27% vs. 64 % in control (p<0.05) and in SFM to 32% vs. 64% in control (p<0.05). The average CP3 level increased at the post-equilibrium stage as compared with control: 33% in TYB group, and 38% in SFM. Similarly, the percentage of CP9 increased: 34% in TYB groups and 26% in SFM. The process of post-thawing increased the percentage of cells with positive intensity of CP3 in TYB group by 75%, in SFM by 73%, and that of CP9 significantly increased in the TYB group by 56% and in SFM by 76%. At the post-equilibrium stage, the following level of active CP3 was detected: in the TYB group 1168± 394.5 vs. 811.3 ± 271.1 in control (p<0.01); and in the SFM group 11314.6 ± 389.4 vs. 811.3 ± 271.1 (p<0.01). The level of active CP9 did not change. However, post-thawing, CP3 activity in positive cells significantly decreased to 580.2 ± 207.6 in TYB and to 727.1 ± 223.4 in SFM (p<0.05). CP9 activity significantly decreased in comparison with control (617.2 ± 135.0) to 344.2 ± 59.0 in TYB group (p<0.05) and 471.7 ± 41.6 in SFM (p<0.05). The assay of SCSA shows that the mean αt value in fresh samples was 180.9 ± 14.3, which significantly decreased in all stages of cryoprocesses to 165.8 ± 20.7 (p<0.05) in TYB and 164.7 ± 16.4 in SFM (p<0.05). The percentage of COMPαt cells in the post-thaw did not differ from the control. ConclusionHuman sperm cells exposed to cryoprotectors increase their apoptotic markers both in quantity and in intensity, however, post-thawing, there was further increase in the percentage of cells exhibiting these markers but their intensity levels decreased. The underlining mechanism of this phenomenon in now investigated. Cryopreservation processes have a tendency to increase the quantity of sperm cells expressing apoptotic markers. On the other hand, at stages of equilibrium and post-thawing, increase in intensity of caspases and chromatin stability was found. We will examine the differences between the cryoprotective media by virtue of this study. Human sperm cells exposed to cryoprotectors increase their apoptotic markers both in quantity and in intensity, however, post-thawing, there was further increase in the percentage of cells exhibiting these markers but their intensity levels decreased. The underlining mechanism of this phenomenon in now investigated. Cryopreservation processes have a tendency to increase the quantity of sperm cells expressing apoptotic markers. On the other hand, at stages of equilibrium and post-thawing, increase in intensity of caspases and chromatin stability was found. We will examine the differences between the cryoprotective media by virtue of this study.