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Showing 119 results for Mice

Fahime Sadat Kamali, Rasoul Shahrooz, Golamreza Najafi, Mazdak Razi,
Volume 0, Issue 12 (12-2019)

Background: Paraquat (PQ), as a pyridine compound, is widely used worldwide to control annual weeds. The oxidative stress caused by PQ can cause deleterious changes in the testicular tissue.
Objective: An investigation on the protective effects of Crocin (CCN) against PQ-induced oxidative damages and apoptotic indices in testicular tissue. 
Materials and Methods: Twenty-eight adult male albino mice (20-25 gr) were divided into four groups (n = 7/each). The control group received 0.1 ml/day of normal saline by intraperitoneal injection (IP); sham-control group received PQ 5 mg/kg/day, IP, and the experimental groups received PQ (CCN+PQ) and CCN-sole (200 mg/kg/day, IP), respectively, for 35 continuous days. At the end of the treatment period, the testes were dissected out and used for biochemical, molecular, and histological analyses. The expressions of tumor suppressor p53, B-cell lymphoma 2 (bcl-2), and caspase-3 were considered as hallmark factors of mitochondria-dependent apoptosis. Moreover, the testicular superoxide dismutase (SOD) and malondialdehyde (MDA) were evaluated as key biomarkers for oxidative stress.
Results: The PQ significantly (p< 0.02, p< 0.01) diminished the spermatogenesis indices and SOD, increased MDA levels, and enhanced the apoptosis-related gene expression. However, the co-administration of CCN and PQ significantly (p< 0.01, p< 0.01, p< 0.02) ameliorated the spermatogenesis ratio, upregulated the SOD level as well as bcl-2 expression, and reduced the MDA content and apoptosis vs the PQ-sole group.
Conclusion: This study showed that the antioxidant properties of CCN enable to ameliorate the PQ-induced destructive effects by upregulating the testicular structure, antioxidant and apoptotic status.
Mina Omidi, Akram Ahangarpour, Seyed Ali Mard, Layasadat Khorsandi,
Volume 0, Issue 12 (12-2019)

Background: Aging is accompanied by decreasing general function in the cells and tissues. D-galactose (D-gal) induces aging and plays a role in the pathogenesis of it. Myricitrin is a plant-derived antioxidant.
Objective: The present study was performed to evaluate the effects of myricitrin on antioxidant defense, sex hormone levels, uterus, and ovarian histology in D-gal-induced aging female mouse model.
Materials and Methods: In this experimental study, 72 female adult NMRI mice, weighing 30-35 gr, 3-4 months old, were randomly divided into six groups (n = 12/each): (I) Control (vehicle; normal saline), (II) D-gal at 500 mg/kg/d for 45 days, (III-V) D-gal + myricitrin-treated groups (these groups received myricitrin at 5, 10, and 20 mg/kg/d, and (VI) D-gal + 100 mg/kg/d vitamin E orally for the last 28 days. The antioxidant indices were done on the basis of colorimetric method, and sex hormone levels were measured by using enzyme-linked immunosorbent assay kits. Histological assessment of the uterus and ovaries were also evaluated.
Results: D-gal impaired the estrous cycle, also degenerative changes occur in the ovarian follicles and damage to the uterus and ovarian tissue occurs. In D-gal group, the level of sex hormones (p =0.03) and the total antioxidant capacity (p = 0.002) decreased, while the level of malondialdehyde and gonadotropins increased (p = 0.03). Myricitrin at lower doses and vitamin E ameliorated the D-gal effects.
Conclusion: These findings suggest that myricitrin at low doses can effectively prevent D-gal-induced oxidation and aging in mice. The effect of myricitrin was equivalent and sometimes better than vitamin E.
Davoud Kianifard, Professor Ali Ehsani, Dr Parisa Zeinolabedini Daneshgar, Dr Ghasem Akbari, Mr Seyyed Maysam Mousavi Sjoar,
Volume 0, Issue 12 (12-2019)

Background: Paclitaxel (PTX), a chemotherapeutic agent, and monosodium glutamate (MSG) have oxidative effects on testicular tissue.
Objective: In this study, the effects of MSG administration on the exacerbation of testicular tissue alterations related to PTX treatment were evaluated.
Materials and Methods: MSG (30 & 60 mg/kg i.p.) was administrated to six groups (n = 8/each) of adult mice before or after PTX treatment: control, PTX-treated, MSG30 + PTX, MSG60 + PTX, PTX + MSG30, and PTX + MSG60. Following the euthanizing, the body weight measurement, pituitary–testicular axis hormonal analysis and serum lipid peroxidation index assessment was prepared, testicular histomorphometry (tubular diameter and germinal epithelium height), immunohistochemistry of p53 was completed. Microscopic indices of spermatogenesis (tubular differentiation, spermiogenesis and repopulation indices) were studied.
Results: Body weight was not changed significantly. The levels of testosterone (p = 0.0001), follicle stimulating hormone (p = 0.019), and luteinizing hormone (p = 0.08) were decreased while the level of lipid peroxidation index was increased (p = 0.208) in the treated groups. The histomorphometry indices (p < 0.0001 and p = 0.001, respectively), germ cells population (p < 0.05) and microscopic indices of spermatogenesis (p = 0.001, p = 0.005, p < 0.0001, respectively) were significantly reduced in all treated groups. The administration of MSG before PTX treatment induces more changes. The most positive reaction to p53 was observed in MSG30 or 60 + PTX groups compared to other groups.
Conclusion: The administration of MSG could intensify testicular tissue alterations related to PTX chemotherapy.
Ali A Movassagh-Pour, Mojdeh Salehnia, Ali A Pourfatollah, Sayed M Moazzeni,
Volume 1, Issue 1 (1-2003)

Background: Embryonic stem cells (ESc) are pluripotent cells which have been used as a model to study 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 stage embryos in mice. Materials and Methods: Blastocyst stage embryos from pregnant NMRI mice were obtained and cultured for 24 h in DMEM medium. 4-6 days after hatching, the inner cell masses (ICM) formed colonies which were then collected mechanically and trypsinized. Several subcultures were prepared in the medium supplemented with 0.1 mM 2 Mercaptoethanol, 1000 U/ml Leukemia Inhibitory Factor (LIF) and 10% Fetal Bovine Serum (FBS). The (ESc) were recognized by alkaline phosphates histochemistry using azo-coupling method. Results: The results demonstrated that a highly pluripotent stem cell line was derived from the blastocyst stage embryos of NMRI mice; however, the rate of colonies was as low as 10%. Conclusion: The LIF is effective to culture and maintain the isolated ICM colonies in undifferentiated condition in the absence of feeder layer.
Mojdeh Salehnia,
Volume 1, Issue 1 (1-2003)

Background: The aim of this study was to determine the correlation between ultrastructural studies for pinopodes expression after ovarian hyperstimulation and progesterone injection in mice. Materials and Methods: Adult NMRI mice were superovulated using human menopasual gonadotropic (hMG) and human chorionic gonadotropic (hCG) hormones; after that, daily injection of progesterone (1 mg/mouse) was performed. Animals were sacrificed by cervical dislocation 3.5 and 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 samples were studied same as experimental groups. Results: 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. In progestrone-injected group, well developed pinopods were expressed 3.5 days after hCG injection and they were transformed to small projections on the fourth day following hCG injection. Also, the life span of pinopods was limited to a short time. At the TEM levels, the pinopods were seen as swelling process on the apical surface, which were more pronounced on day 3.5 of hCG injection in hyperstimulated and progestrone injection. Conclusion: The progestrone may cause premature expression of pinopodes and the implantation failure after ovarian induction may be due to these timing changes.
Behnaz Sheikholslami, Mojdeh Salehnia, Mojtaba Rezazadeh,
Volume 2, Issue 1 (7-2004)

The cytokine of granulocyte macrophage colony stimulating factor (GM-CSF) is a glycoprotein, which is synthesized in the female reproductive tract and has embryonic trophic effect in mammals. The objective of this study was to examine the optimal dosage of GM-CSF to improve the mouse embryo development in vitro. To collect two and eight cells embryos, the pregnant NMRI mice were sacrificed by cervical dislocation at 48 h and 72 h post hCG injections, respectively. The embryos were cultured randomlly in T6 medium supplemented with 5 mg /ml bovine serum albumin (BSA) and 0, 2, and 10 ng / ml human rGM-CSF. The data of blastocyst formation and hatching in different groups of embryo culture were compared by chi-square analysis. The results showed that the developmental rates of 2 and 8 cells embryos to hatching blastocyst in the presence of 2 ng/ml of GM-CSF their control groups (51.5% and 49.7%, respectively) were more than those in the other groups, but insignificant. It seems more researches are necessary to confirm this suggestion that the GM-CSF with 2 ng/ml concentration may have a better potential, not only to enhance the developmental rates of 2 and 8 cells embryos but also for decreasing the degeneration of those embryos.
Mahmoud Hashemitabar, Babak Ghavamizadeh, Fatemea Javadnia, Esmaiel Sadain,
Volume 2, Issue 1 (7-2004)

Background: The luteal phase defect is a common event following the ovarian stimulation. The aim of the present study was to evaluate the use of human chorionic gonadotropine (hCG) and progesterone hormones to improve the luteal phase defect. Materials and Methods: 60 mice were superovulated routinely with human menopausal gonadotropin (hMG) (7.5U) and hCG (10U). The mice were mated and divided into 3 groups: 1- control (n=20) 2- hCG treatment (n= 20), and 3-Progesterone treatment (n=20). Each group was divided again into two subgroups. The mice (10 from each group) had no injection in group one and were injected intraperiteneal (IP) by hCG (5U/day) and progesterone (1mg/day) subcutaneously (sc) in groups 2 and 3, respectively for four days. On the day 5, the animals were killed by cervical dislocation and the uterus were flushed to count the number of blastocyst and their quality. The above treatment were carried out for 12 days in the other 10 mice in each group. Similarly group one had no injection and groups 2 and 3 were injected by hCG and progesterone for 12 days respectively by the same manner as mention above. The animals were killed on day 13 and the implanted embryos were counted. The uterus and ovary were processed on days 5 and 13 of pregnancy for histological studies. Results: The mean number of blastocysts per mouse were: 12.2%, 2.6% and 3% in group 1 to 3, respectively. The nomber of implanted embryos were 29 as: 13 living fetus in one mouse and 16 resorption fetus in the other. The morphology of uterus on day 5 was as follow: no development in the stroma and endometrial gland in control group, the stroma and endometrial gland so developed to form the saw teeth appearance which indicated on receptivity of uterus in hCG treated group similar to progesterone treated group, but without the saw teeth appearance. The continuation of hCG injection maintained the receptivity of uterus; while, the continuation in progesterone caused metaplesia of epithelium. The morphology of ovaries in all three groups showed no changes in corpus luteum size on day 5, and showed the following changes on day 13: increasing the number of primary and secondary follicles in control group; while, reducing the size of corpus luteum in hCG group. Conclusion: Progesterone did not improve the uterus and implantation rate. The prolonged usage of progesterone can change the morphology of uterus to more abnormal state in conterast to the prolonged usage of hCG.
Iraj Rashidi, Mansoureh Movahedin, Taki Tiraihi,
Volume 2, Issue 2 (7-2004)

Background: Pentoxifylline (PX) prevents cAMP breakdown by inhibiting the activity of the cAMP-phosphatase and presumably, stimulates sperm motion. Incubation with PX causes hyperactivation of sperm, an important step in achieving fertilization, and leads to changes in membranes associated with sperm capacitation. Objective: The purpose of this study was to examine the effects of pentoxifylline on sperm viability, motility and fertilization rate after mouse sperm preservation. Materials &amp; Methods: Epididymal spermatozoa from adult NMRI mice were collected in T6 medium supplemented with 5% BSA and divided into four control and four experimental groups. The control groups included: (1) Fresh sperm sample (2) Preserved sperm sample at room temperature for 18 hours. (3) Preserved sperm sample at incubator 37°C for 18 hours. (4) Preserved sperm sample at 4°C for 18 hours. Experimental groups were the same groups after treatment with 3mmol/L PX. All the samples were assessed according to World Health Organization Criteria. Oocytes from superovulated NMRI female mice were inseminated in-vitro incubated sperm of all the control and experimental groups. After insemination and washing, the fertilization rate and cleavage rate were assessed by the presence of two pronucleus (2PN) and 2-cell stage embryos. To study the acrosomal reaction of control and treated spermatozoa transmission electron microscopy (TEM) technique was used. Results: The results showed that addition of 3mmol PX to preserved mouse spermatozoa at 4 ºC and 37 ºC could increase the motility rate significantly (P&amp;lt;0.05) and also it could enhance abnormal morphology rate. Significant increase of fertilization rate was seen after preservation of treated sperm at 4 ºC (P&amp;lt;0.05), but there was not seen significant difference regarding cleavage rate comparing treated and non-treated spermatozoa (P&amp;gt;0.05). Studies with electron microscopy showed that addition of PX to the preserved spermatozoa prevent early acrosomal reaction. Conclusion: The results of this study demonstrated that addition of pentoxifylline in mouse sperm samples after short time preservation can enhance the motility and fertilization rate, although it can enhance the abnormal morphology. It also can increase the number of intact sperm after preservation Article
Mahmoud Hashemi-Tabar, Fatemeh Javadnia, Mahmoud Orazizadeh, Maryam Baazm,
Volume 3, Issue 1 (7-2005)

Background: Recently, embryonic stem (ES) cells have become very important resources in basic medical researches. These cells can differetiate into derivatives of all primary germ layers. Objectives: In order to isolate embryonic stem cells in vitro, the blastocyst were cultured and the morphological aspects, population doubling time, alkalin phosphatse and differentiation properties of the cells were investigated. Materials and Methods: The balstocysts from NMRI mice were cultured for 3 days up to time that inner cell mass (ICM) reach to the outgrowth stage. The cells were disaggregated and trypsinized every 3 days until the appearance of the colonies of ES cells. The colony positive cells were fixed and stained for alkaline phosphatase. The ES cells were cultured in suspension state for 5 days, at the same time Leukaemia Inhibitory Factor (LIF) was removed from media to form embryoid bodies(EBs). The EBs were cultured for 8 - 20 days on collagen coated dish to induce the spontaneouse differentiation. Results: During the 6-9 days after the disaggregation of ICM in the expansion stage, the colony of ES cells appeared as a flat monolayer mass with strike boundaries and nondistinguish cytoplasm including a few nuclei. In colony formation stage, the morphology changed from flat monolayer to round multilayer with strike define boundaries. Undifferentiated cells were seen as intensely small cells attached together compactly with high nucleus/cytoplasm (N/C) ratio. The cells of colonies tend to differetiate by separation from each other and became larger and diffused on substrate by attaching to dish. The positive alkaline phosphatase cells were seen in typical morphology of ES colonies. The EBs cells were seen in culture after 5 days in suspension and began to spontaneously differentiate into various types of cells such as nerve and hematopoitic lineages. Conclusion: Despite strike morphology of ES colonies, it is difficult to distinguish the differentiated from undifferentiated cell colonies in the colony formation stage. New ES cells are capable to give rise into EBs and are susceptible of spontaneously differentiation in various type of cells.
Abbasali Karimpor Malekshah, Amir Esmailnejad Moghaddam,
Volume 3, Issue 2 (7-2005)

Background: In the field of mammalian embryo culture, the putative influence of autocrine/ paracrine factor(s), produce by the embryos itself, is under investigation. A smaller medium drop can prevent dilution of this factor(s). Objective: The objective of this study was to examine the effect of culture medium volume on in vitro development of mouse 2-cell embryos. Materials and Methods: The embryos were obtained from female NMRI mice. To evaluate the effect of medium volume, groups of 16-20 late 2-cell embryos were cultured in 2, 5, 10, 20, 50 and 100 �l of drops of Ham�s F10 medium for 72 h. Results: Development to blastocyst stage in 50 and 100 �l of drop were significantly higher than this in any other volume (p&amp;lt;0.001). Almost a similar pattern was also observed for hatched blastocyst formation. However, the total number of cells in blastocysts, developing in different volumes, were not significantly different. Conclusion: These results indicate that the optimal volumes of Ham�s F10 medium for mouse early embryo development are 50 to 100 �l. However, volumes as small as 2 �l can successfully support mouse 2-cell embryo development to blastocyst and hatching stages.
Mohammad Hossein Dehghan, Thomas Martin, Robabeh Dehghanan,
Volume 3, Issue 2 (7-2005)

Background: About 90% of the world�s contraceptive users are women. This gender-based usage has occurred due to the emphasis of family planning programs and contraception research. Condom, vasectomy and withdrawal are the only male contraception devices available with less assurance for men. For new male contraceptive to have an impact, they must be acceptable to both men and women, as well as effective. A hormonal method will likely come to the market within the next few years. It is necessary to use biologically active botanical substances or fertility-regulating agents of plant origin which are ecofriendly. Objectives: The epididymis is a site which can be exploited for male contraception without undue side effects. It was therefore of interest to investigate the effect of biologically active botanical ecofriendly plants such as Azadirchta Indica (neem) seed alcoholic extract as an efficient and competent male contraceptive on male mouse epididymis. Materials and Methods: In this experimental case control study sixty adult healthy mice divided into two groups of 40 as the control and 20 as the treated group. The treated group was administered by Iranian Botanical Azadirachta Indica seed alcoholic extract, cultivated at Dashteh Moghan (Ardabil province). The seeds was extracted with ethanol then administered first 50 mg/kg body weight /day then 100 mg/kg body weight/day orally for 15 days, following WHO guide lines (MB-50). The target organ, epididymis parameters viz. sperm motility, sperm count fertility rate, Scanning Electron Microscopic (SEM) morphology of spermatozoa and ATPase activity of epididymis of the two groups were compared. Results: The 50 mg/kg body weight (BW)/day showed no significant change in epididymal sperm motility, as compare to the control. Therefore the dose was changed to 100 mg/kg BW/day for 15 days. The body and organ weights (epididymis) of the treated animals were not significantly changed as compare to control group (p&amp;gt;0.05). The treatment brought about a significant reduction in fertility rate when normal cycling female mice were mated with treated males (p&amp;lt;0.001). Decline in ATPase activity in caput and cauda epididymis was observed (p&amp;lt;0.001). SEM photographs showed spermatozoa with abnormal head and bent mid-piece region. Conclusion: Decrease in ATPase activity could be attributed to androgen dependent parameters. However, the fertility rate was also significantly reduced which can be due to the decrease in cauda epididymal sperm motility and their morphological abnormalities. Since the effect on epididymal sperm motility and morphology was manifested in short period of 15 days, it is evident that the extract has potential as an antifertility agent. As this extract do not cause change in the body and organ weight, it is likely that no effect occurred on electrolyte balance
Reza Mahmoudi, Aligholi Subhani, Parichehr Pasbakhsh, Farideh Etesam, Iraj Amiri, Mozhdeh Salehnia, Farid Abolhasani,
Volume 3, Issue 2 (7-2005)

Background: In vitro maturation (IVM) of oocytes is a promising technique to reduce the costs and avert the side-effects of gonadotropin stimulation for in vitro fertilization (IVF). The pregnancy rates from oocytes matured in vitro are much lower than those of in vivo stimulation cycles, indicating that optimization of IVM remains a challenge. Objective: In this study, we investigated the effect of cumulus cells on maturation and fertilization rate of immature oocytes (Germinal vesicle). Materials and Methods: Germinal vesicle (GV) oocytes were recovered from 6-8 weeks old Balb C female mice 48hr after injection of 10 IU pregnant mare serum gonadotropin (PMSG). Collected oocytes were divided into two groups. Group A: GV oocytes without cumulus (denuded oocyte). Group B: GV oocytes with cumulus cells (cumulus-oocyte complex). The oocytes in both groups were cultured in TCM-199 medium in a humidified atmosphere of 5% CO2 in air at 37�C. The maturation, fertilization and developmental rates were recorded after 24hr. Results: Maturation, fertilization and developmental rates in denuded oocytes (DO) were 65.1%, 68.02%, 78.63% respectively, and in cumulus-oocyte complex (COC) were 78.20%, 85.57% and 85.05%, respectively. The maturation, fertilization and developmental rates of COC were significantly higher than those of DO (p&lt;0.05). Conclusion: The results show that cumulus cells have beneficial effects on maturation, fertilization and cleavage rates of mice oocytes.
Homayoon Babaei, Amin Derakhshanfar, Seyed Noureddin Nematollahi-Mahani, Fathemeh Nabipour, Akram Zeraatpisheh,
Volume 3, Issue 2 (7-2005)

Background: Retinoids have been suggested to play a role in oogenesis and oocyte survival. Objective: In the present study the effects of retinol palmitate were investigated on differential follicular counts in response to superovulation as well as follicle quality after vitrification of ovaries. Materials and Methods: Ten, 4 week old female BALB/c mice were randomly assigned to either paraffin (n=5) or retinol palmitate (n=5) administration. Vitamin A administered animals received (i.p.) 250 IU retinol palmitate, dissolved in 0.1 ml of paraffin oil on days one and ten followed by superovulation with 10 IU PMSG. Paraffin administered mice were only treated with 0.1 ml of paraffin oil. The collected left ovaries from both paraffin and vitamin A administered groups were considered as non-vitrified and the collected right ovaries from both treated groups underwent vitrification. Ovaries in the vitrified group were frozen sequentially by placing into two vitrification solutions {VS1: 10% ethylene glycol (EG), 10% DMSO in holding medium (TCM-199 + 20% FBS: HM) and VS2: 20% EG, 20% DMSO in HM}. After warming, recovered ovaries as well as non-vitrified ovaries were serially sectioned and examined histopathologically. Results: The proportion of antral follicles in the non-vitrified ovaries from vitamin A administered mice was statistically higher than the non-vitrified ovaries from paraffin administered group (29.4% vs. 15.6%, respectively; p&amp;lt;0.001). No difference due to retinol palmitate injection was observed for the rate of small follicles between the two non-vitrified groups. The percentage of damaged follicles did not show any significant differences between the two vitrified groups (76% vs. 79%). Conclusions: Our results demonstrate that administration of retinol palmitate may improve the response to superovulation through the shift of follicular growth towards antral follicle development. However, no positive effect of retinol palmitate in the quality of follicles is probable when ovaries are vitrified.
Mojdeh Salehnia, Mitra Arianmanesh, Mandana Beigi,
Volume 4, Issue 1 (7-2006)

Background: The preparation of endometrium for embryo reception is dependent on the ovarian hormones, which are affected by ovarian hyperstimulation procedure.
Objective: The aim of this study was to evaluate the changes in morphometrical indices of endometrium by the daily injection of progesterone after mouse ovarian induction.
Materials and Methods: Adult virgin female mice were selected and divided into control and experimental groups. Experimental groups were superovulated using human menopasual gonadotropic hormone (HMG), and human chorionic gonadotropic hormone (HCG), then they, were subdivided into two groups, which one group was also injected daily by progesterone. All control and hyperstimulated groups were rendered pseudopregnant by cervical stimulation. Three and four days after the HCG injection, the samples of uterine horns were aparted and processed for light microscopic studies.
Results: Our results showed that in the progesterone-injected group, the height of surface and glandular epithelium was decreased on day three (17.6±3.55, 10.02±2.6) and day four (16.9±4.24, 1.6±0.84) respectively, and it had low columnar morphology in comparison with the hyperstimulated and control groups. Also the intercellular spaces of stroma in progesterone-injected group were narrower than these in the other groups.
Conclusion: Ovarian hyperstimulation followed by progesterone injection alter the morphometrical indices of surface and glandular epithelium of endometrium, which could affect on its receptivity.
Mohammad Ali Khalili, Morteza Anvari,
Volume 5, Issue 2 (7-2007)

Background: Research studies on reproductive mechanism of laboratory animals are essential for further advancement of assisted reproductive techniques (ART). One of these studies includes the assessment of in-vitro development of pre-implantation embryos. The objective was to compare the cleavage rates and morphology of in-vivo formed 2 to 8 cell embryos and blastocysts with in-vitro culture of the same embryos for 24 h.
Materials and Methods: 6-8 weeks old female NMRI mice were superovulated with 8IU pregnant mare's serum gonadotropin (PMSG, ip). Two superovulated animals were caged with one male mouse for mating. Mated mice were killed by cervical dislocation at different time intervals to collect a total of 200 (50/ each) 2, 4, 8, and blastocyst embryos from uterine tubes and horns. Following morphological evaluation and cleavage rates, all embryos were incubated in Whittingham's T6 media+5% BSA for 24 h. Following incubation at 37ºC in 5% CO2, the cleavage rates as well as morphological feature of each embryo was re-evaluated and compared with the original embryos.
Results: The best quality embryos collected from uterine tubes were at 2-cells stage, which were reduced when compared with in-vivo developed 4-8 cells embryos. 88% and 52% of 2 and 8 cells embryos were respectively at grade A stage.  28 embryos out of 50 eight-cell embryos were at grades C and D after incubation. Following in vitro culture, the development of 16%, 24%, 24%, and 40% of the 2, 4, 8 cells, and blastocysts were arrested, respectively. Also, only 2 blastocysts (8%) reached the hatching stage which in comparison with in-vivo blstocysts were increased (P>0.05).
Conclusion: In-vitro culture of the in-vivo formed embryos reduced their cleavage rates and morphology, especially at more advanced stages. Therefore, it becomes necessary to improve the in-vitro culture condition and to transfer the embryos at early stage to consequently improve the implantation rates.
Mitra Bakhtiari, Aligholi Sobhani, Mohammad Akbari, Parichehr Pasbakhsh, Mehdi Abbasi, Azim Hedayatpoor, Fardin Amidi , Feridoon Sargolzaei,
Volume 5, Issue 3 (7-2007)

Background: Various approaches have been used in the attempts to improve the quality of frozen–thawed mouse sperms. According to literatures, it seems that hyaluronic acid (HA) has an important role on the permeability and motility of sperms and their interaction with gametes.
Objective: For evaluation of HA supplementation on sperm characteristics and fertilization capability, we investigated the effect of different doses of HA on mouse sperm morphology, motility, vitality and fertilization capability after freezing and thawing.
Materials and Methods: The cauda epididymes was removed from 6 male mice with aseptic method. The sperm samples were frozen in 1.8 ml cryotubes with 18% raffinose and 3% skimmed milk containing cryo-protectant solution. HA at the concentration of 750, 1000 or 1250 µg/ml was supplemented to frozen-thawed sperms. Sperm motility was measured with microscope, and fertilization rate was evaluated after routine IVF by counting the fertilized oocytes. For sperm morphology, papaniclau staining was used while; Eosin B was used for the assessment of sperm viability rate.
Results: HA supplementation (750 µg/ml) improved motility parameters (p < 0.05) and increased the fertility rate (p < 0.05). The effect of 1,000 µg/ml HA was also positive on the sperms. But 1,250 µg/ml HA had negative effect on above mentioned characteristic. On the other hand, none of these doses had any effect on sperm morphology.
Conclusion: The dose of 750 µg/ml of HA has the greatest effect on the motility, vitality and fertility rate of sperms after cryopreservation.

Amaneh Mohammadi Roushandeh, Parichehr Pasbakhsh, Zohreh Alizadeh, Mehryar Habibi Roudkenar,
Volume 5, Issue 5 (7-2007)

Background: Preparation of oocytes is one of the critical factors that determine the developmental competence of embryos produced by in vitro fertilization (IVF).
Objective: In this study, the effect of cysteamine, type of media and glutathione (GSH) level on blastocysts development after in vitro maturation of mouse oocytes were investigated.
Materials and Methods: Premature female mice were primed with pregnant mare stimulating gonadotrophin (PMSG), and germinal vesicle (GV) stage oocytes were obtained 45 hr later. GV oocytes were cultured in presence of 0, 50, 100, 200 and 500 µm cysteamine in TCM199 and MEME media. After IVM, MII oocytes were in vitro fertilized (IVF) and in vitro cultured (IVC) in order to observe embryo development. A group of In Vivo Ovulated (IVO) oocytes after priming with PMSG and HCG also were included in this study. 5,5-Dithio-bis (2nitrobenzoic acid) DTNB-recycling protocol was used for GSH assay.
Results: Rate of IVM and IVF were improved in all oocytes treated with cysteamine in the two medium except 500 µm (81% MII rate in TCM and 64% MII in MEME). Rate of blastocyst in 100 µm cysteamine in TCM1199 and 200 µm in MEME was higher compared to control groups (In TCM 45% and in MEME 35%). In vivo MII and GV oocytes represented the highest and lowest GSH level respectively.
Conclusion: Our results revealed that the media and concentration of cysteamine can affects on IVM, IVF and rate of blastocysts development on dose dependant manner.
Somaye Khosravi Farsani, Reza Mahmoudi, Mir Abbas Abdolvahhabi, Mehdi Abbasi, Fateme Malek, Aligholi Sobhani,
Volume 5, Issue 5 (7-2007)

Background: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimizing.
Objective: The aim of this study was to improve the single step and step-wise vitrification effects on maturing mouse GVBD oocytes by ethylene glycol (EG) in conventional straws.
Materials and Methods: Oocytes with compact cumulus cells were cultured for 3hr in TCM199 supplemented with 10% fetal bovine serum (FBS) in 5% CO2 in air. GVBD oocytes were randomly allocated into three groups. (1) Control (non-vitrified group), (2) exposed to single-step vitrification (contained of EG 20%+0.5M sucrose), (3) exposed to step-wise vitrification (2%, 5%, 10%, 20%EG +0.5M sucrose). In vitrification groups,oocytes were thawed and underwent additional 21 hr maturation. Viability of oocytes and maturation to MII stage were analyzed using inverted microscope and additionally by staining of propidium iodide and Hoechst 33342.
Results: All non-vitrified oocytes were viable after 24 hr; however, viability of vitrified samples in single-step group was significantly lower than that of the step-wise and control Groups. Also, the maturation rate in the step-wise group was significantly higher (p < 0.05) compared to single-step.
Conclusion: These results suggest that step-wise vitrification of  GVBD oocytes as compared to single step vitrification  was better in the rate of  survival and in vitro maturation of oocytes.
Mohamed Youssry, Batuhan Ozmen, Yasser Orief, Khaled Zohni, Safaa Al-Hasani,
Volume 5, Issue 5 (7-2007)

Fertilization involves direct interaction of the sperm and oocyte, fusion of the cell membranes and :::::union::::: of male and female gamete genomes. The completion of this process and subsequent embryo development depend in part on the inherent integrity of the sperm DNA. Sperm genome quality has been emphasized for several years as playing a major role in early embryogenesis. There is clinical evidence showing that human sperm DNA damage may adversely affect reproductive outcomes and that spermatozoa of infertile men possess substantially more DNA damage than do spermatozoa of fertile men. Testing DNA integrity may help selecting spermatozoa with intact DNA or with the least amount of DNA damage for use in assisted reproductive techniques (ARTs). This review will focus on how sperm DNA is organized, what causes sperm DNA damage and what impact this damage may have on reproductive outcome.
Abdolhossein Rezaeian, Seyed Mehdi Kalantar, Safar Farajnia, Mehrdad Soleimani, Abbas Baghi, Abbas Aflatoonian, Sirous Zeinali,
Volume 5, Issue 5 (7-2007)

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