In about 40% of infertile couples, the husband was found to be either the single or a contributing cause of infertility and the systemic management ready for those couples with male infertility are restricted. 135 couples with infertility shared herein; the husbands required in vitro activation for their sperm samples, their wives however; had no fertility problem and were submitted ovarian induction protocol. However; Sperm preparation was done using both, the centrifugation indirect swim-up technique with Global medium and density gradient with ALLGrad solution. While ALLGrad 45% is a ready-to-be-used upper layer. There was an acceleration in seminal parameters after preparation using both various techniques. So, a highly significant increment (P≤0.001) in the percentages of progressive forward motility was in grades (A and B) and grade (A) progressive sperm motility, also a significant enhancement (P≤0.05) in percentage of morphologically normal sperms when applying density gradient mechanism in relation to centrifugation swim-up technique. This study concludes that the use of density gradient mechanism with ALLGrad solution and Global culture medium are more efficient in achieving better percentages of progressive forward motility grades (A and B), grade (A) progressive motility and morphologically normal values of spermatozoa than centrifugation indirect swim-up technique by using Global culture medium. However, it was found there was a non-significant difference when we use both techniques concerning other sperm parameters.
Background: Zygote produce from once a sperm fertilizes an egg cell. Then, the zygote (unicellular) will begin chain of cellular cleavages to produce multicellular mass, its embryo, the differentiated to different tissues and organism. The development of the embryo is called embryogenesis. Coenzyme Q10, is an antioxidant produced in the body. It boosts cellular energy and may enhance the immune system. CoQ10 is present and measurable in seminal fluid, the concentration of CoQ10 directly correlates with both sperm count and motility. It is beneficial in the prevention and treatment a wide range of health problems. Objectives: The present study was aimed to investigate the possibility of using coenzyme Q10 to improvein vitro fertilization (IVF), and early embryonic development (ED) in mice as a model for human being. Methods: Superovulation program was achieved to mature healthy female mice with age 10-12 weeks and weight 24-26 gm. After sacrificing female, oocytes were collected and incubated within CO2 incubator for less than 1 hour. Sperm were collected from vas deference of males. Sperm parameters were assessed after 30 min. ofincubation. Mature oocytes were divided into three groups according to theconcentrations of CoQ10 including G1 (control group; SMART medium only), G2 (treated group; SMART medium enriched with 20 M CoQ10) and G3 (treated group; SMART medium enriched with 40 M CoQ10). IVF technique was performed for 3 groups, and assessment of IVF (%), embryonic development stage (%) and abnormal embryo morphology (%) for each embryo stage. Results: Results of the present study appeared significant increment (P<0.05) in the percentages of IVF for both treated groups as compared to the control groups. Also, significant increase (P<0.0P) in the IVF (%) was observed when using 40 M CoQ10 as compared to 20 M CoQ10. Non significant differences (P>0.05%) in the 8-cells embryo stage were assessed among control and treated groups. Conclusion: From the results of the present study it wasconcluded that the coenzyme Q10 (40µM) enriched to the culture medium improved percentage of in vitro fertilization and no effect on embryonic development.
Background: All men have the natural desire to have their own biological children. Cryopreservation of male gametes is an important aspect of human fertility preservation. With the advancement in assisted reproductive technology, indication for sperm cryopreservation is expanding. Objectives: To evaluate the effect of cryopreservation by using Sperm Freeze TM kit on some sperm parameters for infertile patients. Materials and methods: Ninety five semen samples were taken from male patients. The seminal fluid analysis was performed according to the standard criteria of WHO (1999). The patients were divided into four groups depending on their sperm concentration and motility: Normozoospermic, oligozoospermic, asthenozoospermic and oligoasthenozoospermic patients. Cryopreservation in liquid nitrogen was done by using Sperm Freeze TM kit for the samples and thawing was performed after three months. Results: In general, the post-thawing study revealed a considerable highly significant (P <0.001) decrease in the sperm parameters which include the means of sperm concentration, sperm motility, progressive motility and normal sperm morphology as compared to that of pre-cryopreservation. Whereas the percentage of immotile sperm was highly significant (p<0.05) increase as compared to pre-cryopreservation in all groups except for oligozoospermic patients in which there was a significant (p<0.05) increase. Conclusions: After semen cryopreservation, all parameters were significantly decreased.
Background: Carnitines are widely distributed in nature and their potential health benefits have been popularized. The carnitines exert a substantial antioxidant action. It is often sold as a nutritional supplement. The use of carnitine showed some promise in treatment of male infertility. Objectives: This study was designed to investigate the effects of L-carnitine (LC) administered to pregnant mice on some reproductive parameters of male offspring. Methods: Ninety mature male offspring mice (age: 6 weeks) were divided originally into two groups depending on the pregnant mothers were administered either 0.5 mg/Kg L-carnitine (Treated group) or distilled water (Control group) throughout pregnancy period. Whole body and testicular weight, diameter of seminiferous tubules, thickness of spermatogenic cell layers, levels of serum gonadotropins and sperm parameters were assessed. Results: In the present study, weight of whole body and testes of the male mice offspring group where mothers treated with 0.5 mg/Kg L-carnitine throughout gestation period was significantly increased (P<0.05) as compared to the control group. Further, significant increment (P<0.05) was observed in the thickness of spermatogenic cell layers and diameter of seminiferous tubules for the treated group as compared to the control group. Also, levels of serum follicle stimulating hormone (FSH) and Luteinizing hormone (LH) in the treated group showed significant elevation (P<0.05) as compared to the control group. Sperm parameters for the treated group were enhanced significantly (P<0.05) as compared to the control group. Conclusion: Administration of low dose of L-carnitine to pregnant mice improved reproductive performance of male offspring. Further molecular studies are recommended to investigate the effects of L-carnitine administered to pregnant mice on quality and quantity of acrosomal enzymes for male offspring.
Background: Both clomiphene citrate and tamoxiphene citrate, as an anti estrogen, is the most common prescribed medication for male infertility.In humans, ant estrogens interfere with the normal negative feedback of sex steroids at the hypothalamus and pituitary, thereby increasing endogenous gonadotropin releasing hormone (GnRH) secretion from the hypothalamus and follicle stimulating hormone (FSH) and luteinizing hormone (LH) secretion directly from the pituitary. Objective: The objective of this study was to investigate the effect of clomiphene citrate (CC) and tamoxiphene citrate (TC) administered to male mice on quality of sperm motility and activity. Materials and Methods: Ninety six male mice were divided into two major groups, and every one of these groups was subdivided into three minor groups involving control and two treated groups. In the CC study, 48 mice were administrated orally 0, 0.0125 mg and 0.025 mg as control (G 1), low dose (G 2) and high dose (G 3) for 42 days. While, in the TC study, 48 mice were administrated orally 0, 0.01 mg and 0.02 mg as control (G 1), low dose (G 2) and high dose (G 3) for 42 days. Then, the male mice were sacrificed, both vas deferens were collected and assessment sperm motility and activity. Results: The results of present study appeared that the use high dose of CC or TC significantly increment (P<0.05) sperm motility and activity as compared to control groups. Conclusions: Based on the results of this study, it can be concluded that the administration of high dose of either CC or TC increases sperm motility and activity.
Background: Human growth hormone (HGH) is undisputedly a potent hormone with a wide variety of biological effects. The anabolic actions of human growth hormone have made it attractive as a potential agent for catabolic problems in a wide range of clinical conditions. Objectives: In the present study, we investigated the possible relationship between levels of HGH in serum (S. HGH) and seminal plasma (Sp. HGH) and semen parameters in normozoospermic (NZS) men and infertile patients. Materials and Methods: In total, 66 infertile patients with azoospermia (AZS; no. 52), asthenozoospermia (AsZS; no. 6) and oligoasthenoteratozoospermia (OATZS; no. 8), and normozoospermic men (NZS; no. 12) were included in this study. From each subject, semen and blood samples were taken to assess seminal plasma and sperm parameters and levels of S. HGH and Sp. HGH; respectively. Results: The highest S. HGH was evaluated in AZS patients when compared to NZS and other infertile groups. Significant elevation (P<0.05) in the level of S. HGH was assessed for AZS patients as compared to NZS men. In contrast, the level of Sp. HGH in NZS men was significantly elevated (P<0.05) when compared to AZS patients only. Positive and significant correlations were noticed between levels of Sp. HGH and semen liquefaction time in NZS, AZS and OATZS groups. In NZS men, positive and significant correlation were observed between Sp. HGH and semen volume and negative and significant correlations were detected between Sp. HGH and percentages of sperm motility, progressive motility, immobility and agglutination. Conclusion: Our study represents the first study in Iraq showing that the level of HGH in serum and seminal plasma affect male fertility and sperm parameters in normozoospermic males and infertile patients. Further biochemical and molecular studies are recommended to investigate the mode of action of HGH and its receptors in relation to fertility.
Background: In vitro maturation (IVM) is a culture technology that enables the immature oocytes to reach metaphase II (MII). Vitrification of immature oocytes followed by in vitro maturation can offer advantages, such as to avoid the use of large doses gonadotropins and also be an alternative approach to the use of endocrine stimulation to obtain pre-ovulatory oocytes in all cycles. Objective: The objective of the study is to determine the effect of vitrification technique and several biochemical materials on outcomes of in vitro maturation. Materials and Methods: Normal and viable immature oocytes divided into two major groups including: control group and vitrification technique group. Post-thawing, each major group subdivided into four in vitro maturation (IVM) groups using SMART medium with special additives involving: (G1) group: contained gonadotropins (Gn) only, (G2) group: supplemented with Gn and sucrose (4%), (G3) group: consisted of Gn, sucrose (4%) and follicular fluid (FF) (5%), and (G4) group: contained Gn, sucrose (4%) and FF (10%).Then, assessed the results of in vitro maturation. Results: The results of in vitro maturation for control group shows non significant differences (P>0.05) among all groups. While, a significant increase (P<0.05) in percentage of IVM was achieved when compared G3 and G4 groups with G1 and G2 groups in vitrification technique. Therefore, comparison the results of IVM between control group vitrification technique group appeared a significant difference (P<0.05) in G1 groups and also between G2 groups. However, non significant difference (P>0.05) was appeared between G3 groups and also between G4 groups when comparing between control group and vitrification technique group. Conclusions: Vitrification of immature oocytes were appeared as a promise technique for preservation of oocytes. As well as, the addition of follicular fluid (FF) to maturation medium produced the best results for in vitro maturation (IVM).
Background: Oocyte vitrification is a promising technique. The choice of appropriate types and concentrations of cryoprotectants is essential for the success of the oocytes vitrification. Objective: This study aimed to investigate the effect of vitrification on viability and morphology of oocytes and to compare the effect of several cryoprotectants on the viability and morphology of oocytes during vitrification and post-thawing. Methods: The sheep ovaries were collected from the local abattoir. Only normal and viable oocytes were included in this study. By using cryotop, immature oocytes that were viable with normal morphology were vitrified with 15% DMSO and 15% EG supplemented with 0.0 M, 0.25M, or 0.5 M of either sucrose or trehalose as control and treated groups. Oocytes viability and morphology were assessed post-aspiration and post-thawing. Results: Fromthe results of the present study, the percentage of post-thawing normal and viable oocyte reported with the use of 0.5M trehalose and EG in vitrification solution (VS) significantly)P<0.05) higher than the percentage of post-thawing normal and viable oocyte reported with use of 0.25 M trehalose and EG. Conclusion: Vitrification is simple technique and easy to perform but it needs some experience to prevent any oocyte loss during vitrification and thawing processing. The use of 0.5 M of either sucrose or trehalose in vitrification solution improves the percentage of post-thawing viable and normal oocytes.
Back ground: Optimal IVF and embryo culture conditions depend on the composition of culture media being the most important determinants of successful in vitro interaction of the gametes and subsequent embryo development. ATP is fundamental factor to maintain the life, by providing energy, and controlling the cell function and metabolism. Objective: To investigate the effect ATP supplied to culture medium on fertilization rate and in mice as model for human being. Materials and Methods: The female mice (90) were superovulated using superovulation program (SOP) to produce large number of oocyte were divided into three groups, G1 inseminated with vassal sperm in vitro alone (control) , G2 inseminated with vassal sperm in vitro using SMART enriched with ATP(1.25mM). While, G3 inseminated with vassal sperm in vitro using SMART medium enriched with ATP (2.5mM). Mouse spermatozoa were collected from both vas deferens. Then sperm parameters were assessed after 30 min. IVF technique was performed for 3 groups. Results: The results of the present study appear, significantly increment (P<0.05) in the IVF (%) was observed when using 2.5 mM ATP as compared to 1.25 mM ATP and control group. Conclusions: The result showed that addition of high ATP concentration (2.5 mM) enriched to the culture medium improved percentage of in vitro fertilization.
This study was designed to assess the effect of different concentrations of L-carnitine and Co enzyme Q10 (COQ10) supplied to SMART Pro-medium on human sperm motility and sperm grade activity ( progressive, non-progressive and immotile) during in vitro sperm activation. Eighty seven samples of semen of infertile and fertile men were randomly collected from the subjects at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University. The mean age of the men was (32.034 ± 0.57) years with mean duration of infertility (4.644 ± 0.25) years. Seminal fluid analyses were done involving macroscopic and microscopic examinations according to WHO criteria. Swim-up technique was dependent for in vitro sperm activation , the washed samples were divided before using centrifugation swim-up technique into 3 groups: control group (G1) without L-carnitine and CoQ10. While, in G2, G3 two concentrations of L-carnitine and CoQ10 (20 µg , and 40 µg ) were used. Moreover, all groups within post-activation enriched with SMART-Pro media. The sperm motility and sperm grade activity were evaluated after the addition of L-carnitine and COQ10. The results showed a significant (P<0.05) differences were showed in sperm motility and sperm grade activity of post-activation when compared with the pre-activation. In G2 and G3, with dose of ( 20 µg and 40 µg) of L-carnitine and CoQ10, the results showed a significant (P<0.05) increase in the percentages of sperm motility, progressive sperm motility in spite of the increase in the percentages of sperm motility, progressive sperm motility in G3 compared to G2, there was non significant (P>0.05) differences between G2 and G3. The results for non-progressive sperm motility% and total progressive sperm motility millions/ mL. for 40 µg ( G3) groups were significantly (P< 0.05) decreased when compared to control group (G1) with non-significant (P>0.05) differences between 20µg (G2) group, with significant (P< 0.05) decreased in immotile sperm percentage in post-activation when compared to the pre-activation. From the results of the study, it can be concluded that the addition of 40 µg of L-carnitine and CoQ10 to washed sperms can improve sperm motility and sperm grade activity in vitro. Data were analyzed statistically using complete randomized design (CRD) (one way ANOVA).