Effect of culture, incubation and acrosome reaction of fresh and frozen-thawed ram spermatozoa for in vitro fertilization and intracytoplasmic sperm injection

1997 ◽  
Vol 9 (7) ◽  
pp. 665 ◽  
Author(s):  
M. C. Gómez ◽  
J. W. Catt ◽  
L. Gillan ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Acrosome Reaction ◽  
Vitro Fertilization ◽  
Fertilization Rates ◽  
Frozen Semen ◽  
Acrosome Integrity ◽  
And Control ◽  
Fresh Semen ◽  
Percoll Centrifugation

This study evaluated different sperm treatments for fertilization of sheep oocytes by intracytoplasmic sperm injection (ICSI) and in vitrofertilization (IVF). In Experiment 1, fresh and frozen semen was separated by Percoll centrifugation and incubated at 30°C or 39°C in HSOF or BSOF medium for 1 h before use for IVF or ICSI. For IVF, oocytes were inseminated and incubated with sperm for 30 min, 4 h and 19 h. Sperm were assessed for acrosome integrity after Percoll centrifugation and 1 h incubation, and those used for IVF were assessed after each period of exposure to the oocytes. Fertilization rates after ICSI were higher for fresh than for frozen-thawed sperm and were highest 19 h after IVF with fresh or frozen-thawed sperm in the presence of HSOF at 30°C. In Experiment 2, fresh semen was separated by Percoll centrifugation and incubated for 5 h in HSOF, and the acrosome reaction was induced with lysophosphatidylcholine. Acrosome integrity was then assessed. Fertilization rates after ICSI were similar for acrosome-reacted and control spermatozoa. These results suggest that induction of the acrosome reaction in spermatozoa before ICSI is unnecessary, whereas a capacitating treatment of spermatozoa is required before IVF.

322 EFFECT OF SODIUM NITROPRUSSIDE ON BUFFALO SPERM CAPACITATION IN VITRO

2007 ◽  
Vol 19 (1) ◽  
pp. 276 ◽  
Author(s):  
L. Boccia ◽  
L. Attanasio ◽  
A. De Rosa ◽  
G. Pellerano ◽  
R. Di Palo ◽  
...  
Keyword(s):  
Sodium Nitroprusside ◽  
Acrosome Reaction ◽  
Production Efficiency ◽  
Control Group ◽  
Sperm Capacitation ◽  
Promoting Effect ◽  
Vitro Fertilization ◽  
Domestic Species ◽  
Frozen Semen

The overall in vitro embryo production efficiency in buffalo is hampered by the poor fertilization rate. It is known that the quality of the frozen semen may affect fertilization efficiency. However, it is not possible to rule out that the process of capacitation, required by spermatozoa to acquire the fertilizing ability, is impaired in the in vitro fertilization (IVF) system. Although several agents have been proven to induce sperm capacitation in vitro, heparin treatment is still the most efficient method in most of the domestic species. There is evidence that capacitation is part of an oxidative process and that nitric oxide (NO) acts as a capacitation inducer in human (Herrero et al. 1999 Biol. Reprod. 61, 575–581) and bovine (Rodriguez et al. 2005 Anim. Reprod. Sci. 85, 231–242) spermatozoa. The aim of the present study was to evaluate whether sodium nitroprusside (SNP), a well-known generator of NO in vitro, improves buffalo sperm capacitation in vitro. Frozen–thawed sperm from a bull previously tested for IVF were treated by swim-up in order to select only the motile population. Spermatozoa were incubated in the presence of 0.01 mM heparin (control group) for 1 h (n = 266), 2 h (n = 270), and 3 h (n = 306), and in the presence of 10 �M SNP for 1 h (n = 302), 2 h (n = 286), and 3 h (n = 260). The concentration of SNP was chosen on the basis of a preliminary dose-response trial (0.1 �M, 1 �M, and 10 �M). Following incubation with these agents, sperm were exposed for 15 min to 60 �g mL-1 of lysophosphatidylcholine, an agent known to induce acrosome reaction only on capacitated spermatozoa. Trypan blue was used first to differentiate live from dead spermatozoa and the dried smears were then fixed in 37% formaldehyde and stained with Giemsa for acrosome evaluation by microscopic examination. The proportion of acrosome-reacted spermatozoa in each group was used to assess the efficiency of capacitation under different incubation conditions. Differences between groups were analyzed by chi-squared test. No dead spermatozoa were found in all groups. Following 1-h sperm treatment with either heparin or SNP, the proportion of acrosome-reacted spermatozoa was similar (35.3% vs. 28.5%, respectively). However, extending the incubation time to 2 h, SNP significantly (P < 0.01) increased the incidence of acrosome reaction compared to heparin (60.1% vs. 44.1%, respectively). Analogously, when the sperm treatment was prolonged to 3 h, SNP gave a significantly (P < 0.01) higher percentage of acrosome reaction compared to the control (68.8% vs. 36.6%, respectively). In conclusion, sperm treatment with SNP for either 2 or 3 h significant improved the efficiency of buffalo sperm capacitation in vitro compared with heparin, that is, the capacitating agent currently used in the IVF system. The promoting effect of SNP indirectly indicates that NO acts as a capacitation inducer in buffalo spermatozoa. Finally, these results suggest the need to evaluate the effect of SNP on the fertilizing capability of buffalo spermatozoa in vitro.

scholarly journals In Vitro Fertilization Outcome After Intracytoplasmic Sperm Injection with Fresh and with Frozen-Thawed Epididymal Spermatozoa

KnE Medicine ◽  
2016 ◽  
Vol 1 (1) ◽  
Author(s):  
Hilma Putri Lubis
Keyword(s):  
In Vitro Fertilization ◽  
Intracytoplasmic Sperm Injection ◽  
Oocyte Retrieval ◽  
Epididymal Sperm ◽  
Vitro Fertilization ◽  
Fertilization Rates ◽  
Sperm Aspiration ◽  
Significant Difference ◽  
Epididymal Spermatozoa

<p><strong>Introduction</strong></p><p>Testicular epididymal sperm aspiration (TESA) is one of the method  to retrieve sperm from the testes in men with azoospermia. The aim of the study is to compare the In vitro fertilization (IVF) outcome of intracytoplasmic sperm injection (ICSI)-ET cycles with fresh testicular epididymal spermatozoa obtained on the same day with  oocyte retrieval and with frozen-thawed testicular epididymal spermatozoa.</p><p><strong>Material &amp; Methods</strong></p><p>A retrospective comparative analysis of  patients who underwent fresh TESA and frozen-thawed TESA in ICSI-ET cycles from January 2012 to December 2014 in Halim Fertility Center was done. Fresh testicular epididymal sperm aspiration (fresh TESA) was performed on the same day with oocyte retrieval in 28 cycles and the frozen-thawed testicular epididymal sperm aspiration (frozen-thawed TESA) was used in 30 cycles.  </p><p><strong>Results</strong></p><p>The two groups were comparable in terms of the ages of male and female patients, etiology of infertility and duration of infertility. Fertilization rates in fresh TESA group were 53,5% and in frozen-thawed TESA group, fertilization rates were 50%. There was no statistically significant difference between the groups. Clinical pregnancy rates in fresh TESA group were 35,7%  and in frozen-thawed TESA group, clinical pregnancy rates were 26,7% and statistically there was no significant difference between the groups.</p><p><strong>Conclusion</strong></p>There is no significant difference in the in vitro fertilization outcome of intracytoplasmic sperm injection (ICSI)-ET cycles between fresh TESA and frozen-thawed TESA .

Liquid storage of ram semen in the absence or presence of some antioxidants

1996 ◽  
Vol 8 (6) ◽  
pp. 1013 ◽  
Author(s):  
WM Maxwell ◽  
T Stojanov
Keyword(s):  
Superoxide Dismutase ◽  
Glutathione Peroxidase ◽  
Intrauterine Insemination ◽  
Fertilization Rates ◽  
Liquid Storage ◽  
Semen Storage ◽  
Acrosome Integrity ◽  
Improvement In Survival ◽  
Fresh Semen

The antioxidants superoxide dismutase (SOD), catalase (CAT), cytochrome c (CHc) and glutathione peroxidase (GP) were added at various concentrations to Tris-glucose-yolk diluent (TGY), and their effects on motility, acrosome integrity and fertility of ram spermatozoa were assessed after extension and liquid storage. All the antioxidants improved the motility and acrosome integrity of spermatozoa, and a combination of SOD and CAT had an additive effect on the survival of spermatozoa stored at 5 degrees C but not at 25 degrees C. There was a linear improvement in survival of spermatozoa with increasing dose of antioxidants except for CAT for which doses higher than 200 U mL-1 were toxic. The proportion of oocytes fertilized in vitro declined with time of semen storage (P < 0.001), and was better for semen diluted with TGY containing SOD or CAT than TGY without antioxidants when stored for 7 days (116/246, 47% v. 25/79, 32%; P < 0.05) but not for 14 days (23/174, 13% v. 8/66, 12%). Fertilization rates were unaffected by the presence or absence in the diluent of CHc or GP. The proportions of ewes with fertilized ova and of recovered ova fertilized were better after insemination with semen diluted in TGY containing SOD and CAT than TGY without antioxidants when stored for 14 days (9/18, 50% and 20/40, 50% v. 2/13, 15% and 5/32, 16%; P < 0.05) but not for 7 days (9/20, 45% and 16/48, 33% v. 8/16, 50% and 24/41, 59%). Pregnancy rates were better after intrauterine insemination of ewes with fresh semen than stored semen (11/18, 61% v. 21/75, 28%; P < 0.01), and with semen stored in TGY containing SOD and CAT than in TGY without antioxidants (15/37, 41% v. 6/38, 16%; P < 0.05).

scholarly journals Association between Follicular Fluid Leptin and Serum Insulin Levels in Nonoverweight Women with Polycystic Ovary Syndrome

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
G. Garruti ◽  
R. de Palo ◽  
M. T. Rotelli ◽  
S. Nocera ◽  
I. Totaro ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Polycystic Ovary ◽  
Serum Insulin ◽  
Fertilization Rate ◽  
Serum Leptin ◽  
Vitro Fertilization ◽  
Insulin Levels ◽  
Fertilization Rates ◽  
And Control

Aims. We evaluated the links between leptin and visfatin levels and fertilization rates in nonoverweight (NOW) women with PCOS (NOW-PCOS) from Apulia undergoing in vitro fertilization/embryo transfer (IVF).Materials and Methodology. We recruited 16 NOW women with PCOS (NOW-PCOS) and 10 normally ovulating NOW women (control-NOW). All women underwent IVF. Androgens, 17-β-estradiol (17β-E2), and insulin levels were measured in plasma and/or serum and leptin and visfatin levels were assayed in both serum and follicular fluid (FF-leptin, FF-visfatin).Results. In NOW-PCOS, both serum and FF-leptin were significantly lower than in control-NOW. In NOW-PCOS, significant correlations were found between BMI and serum leptin and insulinemia and FF-leptin. By contrast, in control-NOW, FF-leptin levels were not correlated with insulinemia. Serum visfatin levels were not significantly different in NOW-PCOS and control-NOW, but FF-visfatin levels were 1.6-fold higher, although not significantly, in NOW-PCOS than in control-NOW.Conclusions. Both serum leptin levels and FF-leptin are BMI- and insulin-related in Southern Italian NOW-PCOS from Apulia. In line with other reports showing that FF-leptin levels are predictive of fertilization rates, lower than normal FF-leptin levels in NOW-PCOS may explain their lower fertilization rate and this may be related to the level of insulin and/or insulin resistance.

Intracytoplasmic sperm injection--clinical results from the reproductive medicine unit, Adelaide

1995 ◽  
Vol 7 (2) ◽  
pp. 219 ◽  
Author(s):  
D Payne ◽  
CD Matthews
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Sperm Morphology ◽  
Implantation Rate ◽  
Clinical Results ◽  
Pregnancy Rates ◽  
Vitro Fertilization ◽  
Fertilization Rates ◽  
Percent Normal ◽  
Supernumerary Embryos

The clinical results of 391 cycles of intracytoplasmic sperm injection (ICSI) performed between June 1993 and July 1994 are presented in this report. A total of 4797 oocytes were collected, of which 3792 were injected. Of these, 2603 (69%) fertilized, with normal and three pronuclear fertilization rates of 65% and 4% respectively. About 6% of the oocytes were destroyed while denuding and during ICSI. There were 373 (95%) embryo transfers from which 119 pregnancies arose, giving pregnancy rates of 32% per transfer and 30% per cycle, and an implantation rate of 15% per embryo. Of the pregnancies, 98 (82%) were ongoing. Supernumerary embryos were frozen in 44% of the cycles and 61 subsequent transfers of 130 frozen-thawed embryos produced 11 pregnancies (18%). Only 47 (12%) patients had less than 50% of their oocytes fertilized (mean 31%) after ICSI, and of these, 8 had no fertilization of 13 eggs. Nevertheless, 37 of these 47 patients had an embryo transfer and 9 achieved a pregnancy with an implantation rate of 14% per embryo. The percent normal sperm morphology weakly correlated with percent fertilization (r2 = 0.027, P < 0.02) but not with the implantation rate (r2 = 0.003, P > 0.05). Fifty-nine patients with only occasional motile sperm in the ejaculate and 23 patients in whom epididymal sperm were aspirated were treated. The fertilization rates (66% and 70% respectively) and pregnancy rates per transfer (32% and 24% respectively) were comparable in these two subgroups. The overall ICSI results were also compared with 515 cycles of routine in vitro fertilization (IVF) which were performed at the same time.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Superoxide dismutase affects the viability of thawed European mouflon (Ovis g. musimon) semen and the heterologous fertilization using both IVF and intracytoplasmatic sperm injection

2003 ◽  
Vol 15 (1) ◽  
pp. 19 ◽  
Author(s):  
Fiammetta Berlinguer ◽  
Sergio Ledda ◽  
Irma Rosati ◽  
Luisa Bogliolo ◽  
Giovanni Leoni ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Culture Media ◽  
Fluorescein Isothiocyanate ◽  
Spermatozoa Motility ◽  
Culture Systems ◽  
Fertilization Rates ◽  
Frozen Semen ◽  
Intracytoplasmatic Sperm Injection ◽  
Acrosome Integrity

This study evaluated the effects of superoxide dismutase (SOD) on viability and acrosome integrity of European mouflon spermatozoa after cryopreservation and on the fertilization rates of sheep oocytes after IVF or intracytoplasmatic sperm injection (ICSI). Frozen semen was thawed and washed with synthetic oviduct fluid supplemented with 0.6% bovine serum albumin. After centrifugation, the spermatozoa pellet was split into two culture systems: (i) without SOD; and (ii) in the presence of 1500 IU mL−1 SOD. Sperm viability and acrosome integrity were evaluated simultaneously, immediately after thawing and after 3, 6 and 9 h of culture (5% CO2, 39°C, 90% humidity), by incubating sperm with propidium iodide and fluorescein isothiocyanate-labelled Pisum sativum agglutinin. At the same time, sperm were assessed for motility using a standard scoring system (independent operators’ observation of sperm) that graded degree of motility (i.e. 1 = immotile to 10 = maximum motility, as observed at the moment of thawing). For IVF, frozen–thawed semen derived from the two culture systems was placed in culture together with in vitro-matured sheep oocytes. For ICSI, semen derived from the same culture systems as that for IVF was used, and incubated for 1 h under standard conditions. The results showed a marked difference (P < 0.01) between the percentages of live spermatozoa in medium with SOD and those obtained in medium alone, after 3, 6 and 9 h of culture. The percentages of intact acrosome spermatozoa were higher in medium with SOD after 6 h (P = 0.05) of culture. Spermatozoa motility decreased significantly in SOD containing medium at 3 and 6 h of culture compared with motility in control medium. Fertilization rates were significantly lower in medium with SOD than in medium alone, whereas in the ICSI system fertilization rates were significantly higher in the presence of SOD. The results indicate that the addition of SOD to the culture media enhances the viability rates and the acrosome integrity of cryopreserved mouflon spermatozoa.

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