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PubMed1992

Sperm nuclear chromatin normality: relationship with sperm morphology, sperm-zona pellucida binding, and fertilization rates in vitro.

OBJECTIVE: To study whether the results of tests of sperm chromatin and deoxyribonucleic acid (DNA) normality are related to fertilization rates in vitro. DESIGN: Normal morphology, nuclear maturity determined by acidic aniline blue stain, and DNA normality determined by acridine orange fluorescence of sperm in insemination medium and the number of sperm bound to the zona pellucida (ZP) of the oocytes that had failed to fertilize in vitro were determined. The relationship between sperm test results and fertilization rates were analyzed by logistic regression. SETTING: Samples were obtained from patients undergoing in vitro fertilization (IVF) treatment. RESULTS: The number of sperm bound to the ZP, the percentage of sperm with normal morphology, and the percentage of sperm with normal DNA were the most significant factors related to fertilization rates in vitro. In patients with normal morphology > or = 15% or with > 10 sperm bound per ZP, the percentage of sperm with normal DNA, the number of sperm bound to the ZP, and motility grade were significantly related to IVF rates. CONCLUSION: In patients with normal morphology > or = 15%, failure of fertilization may be because of defects of sperm-ZP binding or abnormal DNA. Assessment of DNA normality of motile sperm in the insemination medium may aid prediction of fertilization rates in addition to normal morphology and sperm-ZP binding.

Acridine Orange↗
PubMed1994

Comparison of Ca2+ responses in human oocytes fertilized by subzonal insemination and by intracytoplasmic sperm injection.

OBJECTIVE: To evaluate the consequences of bypassing the normal interaction between the sperm and oocyte surfaces for the form of Ca2+ responses developing in oocytes at fertilization. DESIGN: Oocytes were fertilized by subzonal insemination (SUZI) (maintaining the normal interaction between cell surfaces of both gametes) or by direct intracytoplasmic sperm injection, and changes in intracellular free Ca2+ concentration were evaluated by confocal laser scanning microscopy after loading oocytes with a fluorescent Ca2+ indicator. SETTING: Private hospital, public research center, and university-based laboratory. PATIENTS, PARTICIPANTS: Patients participating in an assisted reproduction program. INTERVENTIONS: In vitro fertilization, SUZI, intracytoplasmic sperm injection. MAIN OUTCOME MEASURES: Changes in intracellular free Ca2+ concentration. RESULTS: All oocytes fertilized after SUZI showed an oscillatory Ca2+ response introduced by a short initial phase with faster Ca2+ oscillations. In contrast, oocytes fertilized after intracytoplasmic sperm injection did not show a similar change in the oscillation rhythm. In both cases, Ca2+ increases were propagated throughout the ooplasm in a wave-like manner. CONCLUSIONS: The results show that there is a relationship between gamete surface contact and the form of Ca2+ fluxes accompanying fertilization. When the contact between gamete surfaces is skipped by direct sperm injection to the ooplasm, a delayed, truncated Ca2+ response is produced which, however, can maintain the form of Ca2+ waves and Ca2+ oscillations typical of normal fertilization.

Calcium↗
PubMed1995

Sperm morphology and in vitro fertilization outcome: a direct comparison of World Health Organization and strict criteria methodologies.

OBJECTIVE: To perform a direct comparison of two sperm morphology methodologies with regard to IVF outcome. DESIGN: Blinded comparison of two methods of morphology assessment using the same morphology slides. PATIENTS: Data were obtained from 132 couples in a consecutive series of patients undergoing IVF. MAIN OUTCOME MEASURES: Two practical end points were selected for analysis for each couple: the presence of any fertilization and the number of fertilized eggs. Normal traditional morphology was defined as > or = 40% normal forms in a sample and normal strict criteria was defined as > or = 4%. RESULTS: Traditional morphology demonstrated a higher sensitivity and negative predictive value than strict criteria (87% versus 61%, and 68% versus 36%, respectively). Positive predictive value and specificity were also numerically greater but did not reach statistical significance. Abnormal traditional morphology, but not strict criteria, was associated with reduced fertilization even among samples with normal sperm concentration and motility. Samples with normal morphology were associated with a greater number of fertilized eggs per couple than those with abnormal morphology: this difference was 3.2 fertilized eggs for traditional morphology and 1.6 for strict criteria. Overall, for samples with < 40% by traditional morphology only one case yielded more than two fertilized eggs. In contrast, up to five fertilized eggs were noted for the lowest strict criteria scores. CONCLUSIONS: Comparison of traditional morphology and strict criteria with regard to IVF outcome favored traditional morphology in several areas. In particular, low scores were more predictive of poor IVF outcome.

Female↗
PubMed1995

Manipulation of sperm before intracytoplasmic sperm injection improves fertilization rates.

OBJECTIVE: To determine the effect of sperm manipulation before intracytoplasmic sperm injection (ICSI) on fertilization rates. DESIGN: Three methods of sperm manipulation before direct ICSI were compared in two sibling oocyte trials. SETTING: In vitro fertilization unit within a teaching hospital. PATIENTS: Patients undergoing infertility treatment using ICSI. INTERVENTIONS: Oocytes were inseminated by ICSI. MAIN OUTCOME MEASURE: Fertilization rate. RESULTS: In the first trial, a standard manipulation technique gave a 67% fertilization rate compared with 64% with a minimal manipulation technique. In the second trial the standard technique gave a significantly greater fertilization rate compared with no prior manipulation (67% versus 45%). CONCLUSIONS: Manipulation of sperm before direct ICSI is not mandatory for fertilization but significantly improves the fertilization rate.

Cytoplasm↗
PubMed1987

Use of individual human follicles to compare oocyte in vitro fertilization to granulosa cell in vitro luteinization.

Granulosa-lutein (G-L) cells from individual follicles aspirated during cycles of in vitro fertilization-embryo transfer were examined after 3 and 6 days in culture. G-L cells from follicles that contained an oocyte that fertilized in vitro were compared with G-L cells from follicles that contained an oocyte that did not fertilize in vitro. Spent culture media was assayed for progesterone at days 3 and 6 of culture and luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor content of G-L cells was determined at day 6. G-L cell cultures from follicles that contained an oocyte that fertilized in vitro produced significantly more progesterone over 3 and 6 days of culture than those obtained from follicles in which the oocyte did not fertilize. Furthermore, LH/hCG receptor content after 6 days was significantly higher in G-L cells obtained from follicles with fertilized oocytes compared with follicles with unfertilized oocytes. Increased progesterone output and LH/hCG receptor acquisition demonstrate more maturation or "luteinization" by G-L cells aspirated from individual follicles that contain oocytes that fertilized in vitro.

Cells, Cultured↗
PubMed1988

Fertilization of human oocytes by sperm from infertile males after zona pellucida drilling.

Infertile couples who had failed to achieve fertilization of oocytes in previous trials of in vitro fertilization (IVF) were treated by IVF with zona pellucida drilling. Zona drilling entails use of micromanipulation to introduce a gap in the zona pellucida either mechanically or by localized application of a zona solvent from a microneedle. Ten couples were treated, from whom 63 oocytes were recovered for manipulation. Sixteen eggs were denuded of the cumulus oophorus only, and the remaining 47 eggs were subjected to zona drilling. Of the 16 eggs denuded but not drilled, 4 (25%) were fertilized. Of the 47 oocytes drilled, 31 survived (67%) and 10 of the surviving eggs (32%) were fertilized. The polyspermy rate for drilled eggs that fertilized was high (5/10, 50%), and polyspermic eggs were often penetrated by more than two spermatozoa. The remaining five eggs fertilized after drilling were diploid fertilizations, and in three cases cleavage was followed by embryo transfer, although pregnancies were not obtained. These data indicate that zona drilling has the potential for establishing pregnancies in instances where treatment by standard IVF would fail. In addition, results indicate that the block to polyspermy in human eggs occurs at the level of the zona pellucida.

Female↗
PubMed1997

Reinsemination by intracytoplasmic sperm injection of 1-day-old oocytes after complete conventional fertilization failure.

OBJECTIVE: To analyze the efficacy of intracytoplasmic sperm injection (ICSI) for "rescue" of failed conventional insemination cycles. DESIGN: Retrospective clinical study. SETTING: Private infertility clinic. PATIENT(S): Fifty-four couples with non-male factor infertility undergoing routine IVF within our clinic. INTERVENTION(S): Twenty- to 24-hour-old unfertilized mature oocytes from conventional IVF cycles in which fertilization failure was complete were reinseminated by ICSI to attempt late fertilization and rescue of the otherwise failed IVF cycle. MAIN OUTCOME MEASURE(S): Late fertilization, pregnancy, and embryonic implantation. RESULT(S): Intracytoplasmic sperm injection reinsemination of 489 unfertilized mature oocytes caused degeneration in 50 (10.2%) oocytes, gave rise to normal fertilization in 215 (44.0%) of the injected oocytes, and allowed ET in 48 of the 54 cycles in which initial complete fertilization failure had occurred. One hundred sixty-four (76.3%) of 215 late-fertilized oocytes either were used for fresh transfer or were frozen-stored. Eight viable pregnancies resulted, yielding a 14.8% pregnancy rate per initiated cycle. CONCLUSION(S): Barring potential concerns regarding the chromosomal normality of embryos arising from reinsemination, our results suggest that ICSI is a relatively successful means of rescuing conventional IVF cycles in which fertilization fails completely.

Adult↗
PubMed1997

The ratio between carcinoma antigen-125 levels in the seminal plasma and serum may be a marker for fertilization in intracytoplasmic sperm injection.

OBJECTIVE: To assess the relationship between tumor marker carcinoma antigen-125 levels in seminal plasma and serum and fertilization rates in an IVF program, using intracytoplasmic sperm injection (ICSI). DESIGN: A prospective study. SETTING: IVF Unit, Lis Maternity Hospital, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel. PATIENT(S): Twenty-five infertile patients with severe oligo-terato-asthenospermia syndrome and 25 fertile male donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Serum and seminal plasma carcinoma antigen-125 concentrations and fertilization rate per cycle. RESULT(S): In the infertile group, the seminal plasma carcinoma antigen-125 levels ranged from 22.0 to 1,284.0 U/mL (mean level +/- SD, 229.9 +/- 274.2 U/mL). In the normospermic fertile male donors, the seminal plasma carcinoma antigen-125 concentrations ranged from 12.2 to 336.7 U/mL (mean level +/- SD, 110.1 +/- 91.6 U/mL). This difference was statistically significant. The mean +/- SD ratio between the seminal plasma/serum carcinoma antigen-125 levels differed significantly between the infertile group (47.9 +/- 61.3) and the fertile male donors (5.7 +/- 3.5). In the infertile group, the ratio between the seminal plasma/serum carcinoma antigen-125 levels was found to be negatively correlated with the oocyte fertilization rate. CONCLUSION(S): The ratio between carcinoma antigen-125 levels in the seminal plasma and serum may be an indirect marker for male infertility and fertilization rate in IVF treatment using ICSI.

Adult↗
PubMed1998

Sperm morphology evaluated by computer (IVOS) cannot predict the fertilization rate in vitro after intracytoplasmic sperm injection.

OBJECTIVE: To evaluate sperm morphology assessment using the IVOS (Hamilton-Thorne Research Version 3 Dimension Program, Beverly, MA) system in prediction of fertilization rate in vitro after intracytoplasmic sperm injection (ICSI). DESIGN: A prospective clinical study. SETTING: Diagnostic andrology laboratory and assisted conception service. PATIENT(S): Thirty-five patients from the ICSI program were evaluated. Semen samples were analyzed using a computerized system for conventional semen parameters, sperm movement characteristics, and sperm morphology. Only patients with three or more metaphase II (MII) oocytes available were studied. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Fertilization rates in vitro after ICSI were compared according to the sperm morphology obtained from the IVOS system. RESULT(S): Linear regression analysis of fertilization rates against the sperm parameters assessed by computer (IVOS), which included conventional semen parameters, sperm movement characteristics, percentage of normal sperm morphology, and percentage of each specific abnormal sperm morphology, did not reveal any significant correlations. The mean (+/- SEM) fertilization rates in the healthy prognosis group (normal sperm morphology > or = 4%) and poor prognosis group (normal sperm morphology < or = 4%) were 82.4 +/- 4.0% and 75.0 +/- 3.8%, respectively. There was no statistically significant difference in the mean fertilization rate between both groups. Moreover, no statistically significant difference was found in overall fertilization rates in vitro between the two prognosis categories (79.6% versus 78.0%). CONCLUSION(S): Sperm morphology obtained from the IVOS system is not related to the outcome of ICSI and cannot be used for prediction of fertilization rate in vitro after ICSI.

Adult↗
PubMed1997

Fertilization and pregnancy rates after intracytoplasmic sperm injection using ejaculate semen and surgically retrieved sperm.

OBJECTIVE: To compare the fertilization rates and pregnancy rates (PRs) in intracytoplasmic sperm injection (ICSI) using sperm from ejaculates of normal and abnormal semen, epididymal sperm, and testicular sperm of obstructive and nonobstructive azoospermic patients. DESIGN: Retrospective study. SETTING: The Egyptian IVF-ET Center. PATIENT(S): Three hundred fifty patients underwent 366 ICSI cycles. INTERVENTION(S): ICSI, epididymal sperm aspiration, and testicular biopsy. MAIN OUTCOME MEASURE(S): Fertilization rates and PRs. RESULT(S): Patients were divided into five groups according to the quality and source of sperm. Patients in group 1 underwent 102 cycles of ICSI using ejaculated abnormal semen, group 2 underwent 44 cycles using epididymal sperm, group 3 underwent 82 cycles using testicular sperm from obstructive azoospermia, group 4 underwent 80 cycles using testicular sperm from nonobstructive azoospermia, and group 5 underwent 58 cycles using normal semen. There was no significant difference in the fertilization rates and PRs among groups 1, 2, and 3. In group 4, the fertilization rate and PR were significantly lower than in all other groups. In group 5, the fertilization rate was significantly higher than in all other groups. CONCLUSION(S): The fertilizing ability of sperm in ICSI is highest with normal semen and lowest with sperm extracted from a testicular biopsy in nonobstructive azoospermia. There was no significant difference in fertilization rates and PRs between ejaculated sperm of different parameters and surgically retrieved sperm in obstructive azoospermia.

Adult↗
PubMed1997

The role of in vitro fertilization and intracytoplasmic sperm injection in couples with unexplained infertility after failed intrauterine insemination.

OBJECTIVE: To determine an optimal insemination technique in patients undergoing IVF after failed IUI and the role of intracytoplasmic sperm injection (ICSI) in such cases. DESIGN: Prospective, randomized study in couples with unexplained infertility (n = 63) and mild endometriosis (n = 7) undergoing IVF after four IUI cycles. Sibling oocytes were randomized into standard IVF or ICSI insemination according to the order of retrieval. SETTING: In vitro fertilization program at the Instituto Valenciano de Infertilidad, Valencia, Italy. PATIENT(S): Seventy couples with unexplained infertility undergoing IVF after failing to conceive with controlled ovarian stimulation and IUI. INTERVENTION(S): In vitro fertilization and ICSI. MAIN OUTCOME MEASURE(S): Fertilization, cleavage, and embryo quality were compared in IVF- and ICSI-inseminated oocytes. RESULT(S): There was no significant difference in fertilization rates between ICSI (60.4%) and conventional IVF (54.0%). Similarly, there was no difference in embryo quality between both groups. There was no total fertilization failure in ICSI-inseminated oocytes, whereas 8 (11.4%) of 70 cases showed absence of fertilization when conventional IVF was used. CONCLUSION(S): Couples with unexplained infertility and mild endometriosis failing to conceive with IUI and undergoing IVF have an 11.4% chance of fertilization failure that can be overcome easily by using ICSI in at least some oocytes. ICSI, however, is not superior to IVF as an insemination technique in most cases. These data should be used in counseling patients.

Adult↗
PubMed1998

Validity and cost-effectiveness of antisperm antibody testing before in vitro fertilization.

OBJECTIVE: To determine the usefulness of and cost-effectiveness of antisperm antibody testing in the prediction of poor fertilization rates in couples undergoing IVF. DESIGN: Retrospective cohort study. SETTING: A hospital-based reproductive endocrinology and infertility practice. PATIENT(S): Male partners of 251 couples undergoing IVF between 1992 and 1997. MAIN OUTCOME MEASURE(S): Fertilization rates in couples undergoing conventional IVF. RESULT(S): One hundred nineteen couples were evaluated for antisperm antibodies; fertilization rates were similar in those couples whose husbands were and were not tested (64% versus 68%). Antisperm antibodies were detected in 16 men. Four (25%) of the 16 couples whose husbands had antisperm antibodies fertilized < or = 50% of oocytes, compared with 31 (30%) of the 103 couples whose husbands did not have these antibodies. Overall, 21 couples (8.4%) experienced complete fertilization failure. In a program that included antisperm antibody testing for selected couples and intracytoplasmic sperm injection (ICSI) for those who tested positive, it would cost $11,735 to prevent a fertilization failure (assuming ICSI were 100% effective), whereas it would cost $9,250 to perform ICSI in a second IVF cycle for those who initially failed. CONCLUSION(S): In this practice setting, antisperm antibody testing has low sensitivity in predicting low or no fertilization and does not appear to be cost-effective when selectively ordered as part of an IVF workup.

Adult↗
PubMed1998

Human sperm head morphometric distribution and its influence on human fertility.

OBJECTIVE: To study the distribution of live sperm head size in semen and sperm preparations as a predictor of fertility. DESIGN: Prospective blind clinical trial. SETTING: Academic tertiary referral center. PATIENT(S): One hundred fifty-five patients undergoing IVF treatment. Females with conditions negatively influencing fertilization were excluded. INTERVENTION(S): Morphometric analysis (head area, major axis, minor axis, and elongation ratio) of video images of sperm in semen and swim-up preparations used for IVF treatment was performed with a Hamilton-Thorne analyzer V 8.1 (Hamilton-Thorn Research, Beverly, MA). MAIN OUTCOME MEASURE(S): Oocyte fertilization. RESULT(S): Seventy-four percent of patients achieved fertilization. Fertilizers and nonfertilizers had different sperm head area distribution. The fertilizers had a significantly smaller interquartile range of sperm head area and of major axis in both semen and sperm preparation compared with the nonfertilizers. A subgroup of men who had fathered a child naturally had a more uniform sperm head area in semen with a significantly smaller median compared with those who failed to father a child naturally with their healthy female partner. We used multiple logistic regression applying forward stepwise selection of variables in building three predictive models of probability of fertilization. CONCLUSION(S): Successful IVF or history of fathering a child was associated with a more uniform sperm head area in semen and sperm preparation.

Adult↗
PubMed1983

Sequence-specific adenylations and deadenylations accompany changes in the translation of maternal messenger RNA after fertilization of Spisula oocytes.

A dramatic change in the pattern of protein synthesis occurs within ten minutes after fertilization of Spisula oocytes. This change is regulated entirely at the translational level. We have used DNA clones complementary to five translationally regulated messenger RNAs to follow shifts in mRNA utilization at fertilization and to characterize alterations in mRNA structure that accompany switches in translational activity in vivo. Four of the mRNAs studied are translationally inactive in the oocyte. After fertilization two of these mRNAs are completely recruited onto polysomes, and two are partially recruited. All four of these mRNAs have very short poly(A) tracts in the oocyte; after fertilization the poly(A) tails lengthen considerably. In contrast, a fifth mRNA, that encoding alpha-tubulin mRNA, is translated very efficiently in the oocyte and is rapidly lost from polysomes after fertilization. Essentially all alpha-tubulin mRNA in the oocyte is poly(A)+ and a large portion of this mRNA undergoes complete deadenylation after fertilization. These results reveal a striking relationship between changes in adenylation and translational activity in vivo. This correlation is not perfect, however. Evidence for and against a direct role for polyadenylation in regulating these translational changes is discussed. Changes in poly(A) tails are the only alterations in mRNA sizes that we have been able to detect. This indicates that, at least for the mRNAs studied here, translational activation is not due to extensive processing of larger translationally incompetent precursors. We have also isolated several complementary DNA clones to RNAs encoded by the mitochondrial genome. Surprisingly, the poly(A) tracts of at least two of the mitochondrial RNAs also lengthen in response to fertilization.

Animals↗
PubMed1997

Surgical treatment of testicular trauma: effects on fertility and testicular histology.

Unilateral testicular trauma in the postpubertal male can lead to alterations in semen analysis, but it is not clear what effect this has on fertility. To better understand how surgical treatment of testicular trauma affects both fertility and testicular histology the following study was performed. Eighty postpubertal Lewis rats were divided into eight equal groups with one group serving as a control. In the 70 remaining rats the left testicle was subjected to blunt or penetrating injury. The testicles were either left untreated, were removed, or were repaired with sutures or mesh before being returned to the scrotum. Following recovery, each male was allowed to mate to determine fertility. Fertility rates were significantly lower in all postinjury groups except the postinjury orchiectomy group. Histological analyses showed nonspecific inflammation, smaller tubules, and impaired spermatogenesis in all postinjury testicles regardless of the type of treatment. Contralateral testicles had no evidence of autoimmune injury and were essentially identical to the control group. In the postpubertal Lewis rat, unilateral testicular trauma leads to impaired fertility unless the injured testicle is removed soon after the injury. Various methods of repairing the injury did not improve fertility. In spite of the impaired fertility, the contralateral testicle appears histologically normal.

Animals↗
PubMed2001

Effect of sperm number and oxygenation state of the storage media on in vitro fertility of bovine sperm stored at ambient temperature.

The effects of storage time and the oxygenation state of the storage medium on motility, viability, and in vitro fertility of stored diluted sperm were investigated. Oocytes collected from abattoir material were matured and fertilized in vitro on defined days with sperm stored for up to 11 days in a citrate-based commercial diluent. The proportions of oocytes fertilized and developing to the blastocyst stage were used to assess the quality of the stored semen. In vitro fertility of sperm declined with storage time. There was no significant effect of the oxygenation state of the medium on in vitro fertility of stored sperm. Increased sperm-to-oocyte ratios resulted in a significant elevation of the proportion of oocytes fertilized on day 0 of storage and the proportion of fertilized oocytes developing to the blastocyst stage on days 0 and 3 of storage, suggesting some form of sperm competition or egg selection of sperm based on the ability of sperm to induce normal development.

Animals↗
PubMed2001

Effect of sperm cryopreservation and treatment with calcium ionophore or heparin on in vitro fertilization of horse oocytes.

Little information is available on methods of sperm capacitation for IVF in the horse. In this study, we summarized results of several independent trials that compared acrosome reaction, hyperactivation and chromatin integrity of fresh or cryopreserved stallion spermatozoa after treatment with heparin or with calcium ionophore. We also examined the influence of spermatozoa storage (fresh vs. cryopreserved), capacitation treatment, oocyte maturation time and cumulus morphology on the penetration rate and fertilization rate. We recovered cumulus-oocyte-complexes (COCs) from ovaries by ultrasound guided follicle aspiration or by scraping of follicles from ovaries obtained at a slaughterhouse. Upon recovery, we evaluated the cumulus morphology, and the COCs were matured in vitro for 18 to 24 or 26 to 40 h. Fresh semen and cryopreserved semen were treated either with heparin (200 microg/mL) or calcium ionophore (7.14 microM). Overall, 28.4% (99/349) of the oocytes were penetrated, and 12.9% (45/349) were fertilized. Fresh spermatozoa treated with calcium ionophore showed a higher penetration rate than cryopreserved spermatozoa (36.0 vs. 0%). Fresh and heparin-treated spermatozoa showed a penetration rate of 29.1%, and the same treatment for cryopreserved spermatozoa showed a penetration rate of 33.7%; none of these differences was significant (P>0.05). Fertilization rates after the calcium and heparin treatment followed the same trend and also showed no significant differences. Prolonged maturation period resulted in higher penetration (P<0.05) and fertilization rates in compact (26 to 40 h: 37.7 and 13.1% vs. 18 to 24 h: 13.1 and 2.8%) and in tendency in expanded COCs (26 to 40 h: 40.0 and 30.3% vs. 18 to 24 h: 29.4 and 13.5%). In oocytes with only a few cumulus cells, the rates tended to be higher after the shorter incubation (18 to 24 h: 33.5 and 18.8% vs. 26 to 40 h: 17.2 and 6.5%). We observed hyperactivation more frequently in fresh than in cryopreserved semen after different treatments (43.2, 39.1 and 35.4% for heparin, calcium ionophore and control vs. 15.7, 10.8 and 5.7%, respectively). We observed significant changes in the acrosome reaction of fresh spermatozoa after heparin treatment (62.6 vs. 48.2%, P<0.05), as well as in cryopreserved spermatozoa after calcium ionophore treatment (31.7 vs. 17.6%, P<0.05). The chromatin integrity was significantly reduced after heparin treatment of fresh spermatozoa, in comparison to control and calcium ionophore (81.0 vs. 87.3 and 86.6, P<0.02). We also observed a similar reduction of chromatin quality after heparin treatment in cryopreserved spermatozoa, but the difference was significant only between heparin and calcium ionophore treatment [77.4 vs. 86.4 (P<0.02) and 84.9]. The results in the this retrospective study show that capacitating fresh spermatozoa with calcium ionophore, or using heparin in cryopreserved spermatozoa, results in higher penetration and fertilization rates of in vitro matured horse oocytes. A prolonged maturation time of 26 to 40 h is necessary for compact cumulus oocyte complexes to achieve the fertilization capacity. Further investigation is needed to show the developmental capacity of these fertilized oocytes.

Acrosome Reaction↗
PubMed2003

In vitro fertility and motility characteristics of frozen-thawed boar epididymal spermatozoa separated by Percoll.

Frozen-thawed epididymal spermatozoa from four boars were separated through a Percoll gradient, and motility characteristics and in vitro fertility were assessed. Percoll-separated spermatozoa had a significantly higher percentage of motile and progressively motile spermatozoa than those that were not separated (P < 0.0001). However, there were no clear differences in other motility parameters between Percoll-separated and un-separated spermatozoa. Furthermore, sperm agglutination was decreased by Percoll separation (P < 0.05). The effects of Percoll separation on in vitro fertility of spermatozoa differed among boars. In addition, there were large differences in fertility between sperm samples in vitro. Sperm samples, which indicate highly motile and progressively motile, did not always show high in vitro fertility. Furthermore, there was no distinct pattern between fertility in vitro and motility parameters. There was no difference in fertility in vitro between Percoll-separated and un-separated spermatozoa from two of the four boars. However, in vitro fertility of Percoll-separated spermatozoa was higher than that of un-separated spermatozoa from the other two boars.

Animals↗