Pretreatment of sperm with human follicular fluid for borderline male infertility

To test the usefulness of human follicular fluid (FF) in treating male infertility, we incubated washed sperm specimens from 31 couples undergoing intrauterine insemination (IUI), for male and/or unexplained infertility, with either FF or Ham's F-10 medium (Gibco, Grand Island, NY), in alternating cycles in a randomized manner. Semen specimens from 28 men were incubated with either medium or FF. Incubations with FF have increased sperm penetration assay (SPA) scores from 24.8 +/- 17.3 to 34.3 +/- 13.6 (P less than 0.01). Incubation with heat-inactivized FF also has increased SPA scores, although to a lesser extent than noninactivized FF. Seventeen pregnancies occurred in the 31 couples treated by IUI (54.8%), 16 of them in FF-treated cycles (51.6%) and one in "control" IUI cycles (3.2%, P less than 0.01). All pregnancies occurred within four treatment cycles. Thus, IUI after sperm wash and preincubation with FF may be suggested for four to six cycles to couples with male factor and/or unexplained infertility who are reluctant to resort to artificial insemination by donor or adoption, before attempting the more costly and complex in vitro fertilization-embryo transfer procedure.     

Intrauterine insemination-ready versus conventional semen cryopreservation for donor insemination: a comparison of retrospective results and a prospective, randomized trial.

OBJECTIVE: To compare fecundity rates following intrauterine insemination (IUI) with donor sperm frozen conventionally versus an IUI-ready preparation. DESIGN: Both retrospective results and a prospective, randomized study where recipients were assigned to one of two sperm cryopreservation methods in each cycle of intrauterine insemination are reported. SETTING: University-based infertility practice, affiliated private practices, and andrology laboratory. PATIENT(S): Women desiring therapeutic insemination in an effort to establish pregnancy. INTERVENTION(S): Intrauterine insemination with donor sperm frozen conventionally or by an IUI-ready protocol. MAIN OUTCOME MEASURE(S): Cycle fecundity in donor IUI recipients. RESULT(S): In a retrospective analysis involving 642 inseminations in 209 recipients, 79 pregnancies were recorded for an overall pregnancy rate of 12.3% per insemination (or cycle): 11.3% with IUI-ready sperm and 13.9% with conventionally preserved sperm. In a follow-up prospective, randomized study, the pregnancy rate for IUI-ready sperm preparations was 36% per cycle (14 of 39) whereas that for conventionally preserved sperm was 19.6% per cycle (9 of 46). Thirteen of the 23 pregnancies occurred in the first study cycle of insemination; only two pregnancies were observed in patients undergoing more than four cycles of insemination. CONCLUSION(S): Cycle fecundity for IUI-ready donor sperm is equivalent to conventional cryopreserved sperm based on both prospective and retrospective assessments.     

Comparison of intrauterine and intracervical insemination with frozen donor sperm: a meta-analysis

OBJECTIVE: To determine whether artificial insemination with frozen donor sperm yielded a higher pregnancy rate per cycle by intracervical (ICI) or intrauterine (IUI) techniques. A meta-analysis was performed. DATA IDENTIFICATION: A computerized MEDLINE search of the English-language literature on artificial insemination with donor sperm was performed and augmented by a review of meeting abstract books and references in published papers. STUDY SELECTION: Only prospective randomized studies that reported monthly fecundity rates for IUI and ICI with frozen donor sperm were included. DATA ANALYSIS: Seven studies were identified. The odds ratios (OR) and 95% confidence intervals (CI) were determined with use of the general estimating equation method for the three studies for which raw data could be obtained. For the remaining four studies, the OR and CI were assessed with use of the published summary data. A random-effects meta-analysis was then performed. RESULT: Intrauterine insemination resulted in a significantly higher monthly fecundity rate with a common OR of 2.4 (CI 1.5-3.8). CONCLUSION: On the basis of this meta-analysis of the seven prospective studies, IUI results in higher pregnancy rates than ICI for frozen donor insemination.     

A prospective, randomized study of pregnancy rates after transuterotubal and intrauterine insemination.

OBJECTIVE: To assess the relative efficacy, in terms of clinical pregnancy rates (PRs), of transuterotubal insemination versus the more traditional intrauterine insemination (IUI) procedure. DESIGN: Prospective, randomized, cross-over. SETTING: University-affiliated tertiary care center. PATIENTS: One hundred sixty infertile patients underwent 414 inseminations with or without controlled ovarian hyperstimulation. INTERVENTIONS: All patients were randomized in their initial cycle to transuterotubal insemination or IUI then crossed-over in subsequent cycles (n = 191 total cycles of transuterotubal insemination and n = 223 total cycles of IUI). Transuterotubal insemination was performed initially with ultrasound guidance, and then a tactile technique was used for the last 6 months of the study. MAIN OUTCOME MEASURES: Clinical PRs and complications after both insemination methods. RESULTS: The clinical PR per treatment cycle was 7% (13/191) after transuterotubal insemination and 7% (16/223) after IUI. The overall PR per patient was 18% (29/160). The incidence of ectopic pregnancy was 1 in 191 for transuterotubal insemination cycles and 0 in 223 for IUI cycles. Other complications included 3 vasovagal episodes with transuterotubal insemination and 1 with IUI. There was no clinical evidence of tubal infection, trauma, or perforation in either group. CONCLUSION: Transuterotubal insemination did not appear to be associated with a higher PR when compared with IUI in this study. The potential for increased risk from complications related to the more invasive tubal technique does not appear to justify its use presently.     

A comparison of intrauterine versus intracervical insemination in fertile single women

OBJECTIVE: To compare the efficacy of intrauterine insemination (IUI) and intracervical insemination (ICI) when used by fertile single women in a donor insemination program. DESIGN: Prospective randomized crossover study. SETTING: Donor insemination program (not an infertility clinic). PATIENT(S): Single fertile women choosing to inseminate with frozen donor semen. INTERVENTION(S): Clients received procreative counseling and screening and were then randomly assigned to begin office insemination with ICI or IUI. If additional insemination cycles were required, the clients used the method opposite their previous method of insemination until pregnancy was achieved. MAIN OUTCOME MEASURE(S): Monthly fecundity rate was compared between the two methods of insemination. RESULT(S): Sixty-two women contributed a total of 189 cycles, 94 by IUI and 95 by ICI. The monthly fecundity rate for IUI was 15%, as compared with 9% for ICI, (P=0.14). When the analysis was confined to cycles in which only one insemination was performed (64 IUI and 65 ICI cycles), the monthly fecundity rates were 14% for IUI and 5% for ICI (P=0.04). CONCLUSION(S): Intrauterine insemination with frozen donor sperm is more effective than intracervical insemination for single women without known fertility problems.     

Successful Pregnancy with Sperm Retrieved from the Bladder after the Introduction of a Low-Electrolyte Solution for Retrograde Ejaculation

Objective: To determine the safety and efficacy of the introduction of a low-electrolyte solution into the bladder before masturbation for sperm recovery in patients with retrograde ejaculation.

Design: Two case reports.

Setting: Assisted reproduction practice at a university hospital.

Patient(s): Two men with retrograde ejaculation.

Intervention(s): A 0.32-mol/L glucose solution or Ham’s modified F-10 medium (GIBCO, Grand Island, NY) with 10% serum was introduced into the urinary bladder before masturbation. The retrieved sperm were used for IUI.

Main Outcome Measure(s): The motility of retrieved sperm, achievement of pregnancy, and outcome of pregnancy.

Result(s): In patient 1, the motility rate of sperm retrieved from the bladder after the introduction of Ham’s modified F-10 medium was higher than that after the introduction of a glucose solution. The patient’s wife did not become pregnant during 10 IUI cycles after treatment with Ham’s modified F-10 medium. However, she became pregnant during the first IUI cycle after treatment with a glucose solution and was delivered of healthy twin infants. The wife of patient 2 also became pregnant during the first IUI cycle after treatment with a glucose solution and was delivered of a healthy female infant.

Conclusion(s): The introduction of a glucose solution appears to be a safe and simple method for sperm recovery in patients with retrograde ejaculation.     

Good efficiency of intrauterine insemination programme for serodiscordant couples with HIV-1 infected male partner: A retrospective comparative study

Objective

Assisted reproduction techniques can minimize the risk of HIV female contamination when the male partner is HIV-infected. The aim of this study was to investigate the efficiency of sperm washing and intrauterine insemination (IUI) in these couples.

Study design

Retrospective comparative study. Eighty-four HIV-1 serodicordant couples underwent 294 IUI. The control group was composed of 90 couples (320 IUI cycles) with donor sperm. Spermatozoa from HIV-1 infected male partner were prepared and tested for HIV-1 according to sperm washing method. Spermatozoa from HIV-1 and donor male were frozen before IUI. IUI were performed after ovarian stimulation. Main outcomes measures were pregnancy rate per cycle and baby take-home rate per couples.

Results

Although the pregnancy rate and baby take-home rate were higher in IUI with sperm washing than in IUI using donor sperm (18.0 versus 14.7 and 52.4 versus 41.1, respectively), the differences were not statistically significant. In serodiscordant couples, blood estradiol levels under ovarian stimulation and total motile sperm inseminated were a determining factor in achieving pregnancy. No female HIV-1 contamination occurred.

Conclusion

This study demonstrates that sperm washing and IUI are highly effective in enabling serodiscordant couples with an HIV-1 infected male partner to have a child.     

Quinn's advantage fertilization medium enhances zona pellucida-induced acrosome reaction compared with human tubal fluid medium

The aim of this study was to compare sperm velocity, hyperactivation, zona pellucida (ZP) binding and ZP-induced acrosome reaction (AR) between Quinn's advantage fertilization (QAF), human tubal fluid (HTF) and Ham's F10 media. Semen samples were obtained from normozoospermic men and motile spermatozoa were prepared by gradient centrifugation (PureSperm). Unfertilized oocytes from clinical IVF were used for spermatozoa-oocyte interaction tests. Sperm velocity and hyperactivation were assessed using a Hamilton-Thorn motility analyser. When media were supplemented with human albumin, sperm motility and velocity and sperm binding were not significantly different between QAF and HTF. However, ZP-induced AR was significantly higher with QAF than HTF (42±22 versus 21±18, P<0.th001). Sperm velocity, hyperactivation and sperm binding were also significantly higher in QAF than Ham's F10 media. Supplementation of media with either human serum or human albumin showed no difference in effect on all sperm test results. In conclusion, QAF medium significantly enhances ZP-induced AR which is essential for sperm penetration. Thus QAF appears to be a better medium than HTF for sperm fertilizing ability in conventional IVF.     

Human in vitro fertilization and embryo transfer at the University of Texas,Houston

The current clinical and laboratory protocols of the in vitro fertilization program at the University of Texas at Houston are described and detailed. Most patients treated have tubal disease, with a limited number of couples having male factors. Clomiphene citrate (50 mg/day, cycle days 5–9) with or without added menopausal gonodotropins is the principle agent for enhanced follicular recruitment. Follicular development is monitored primarily with daily ultrasonography, and human chorionic gonadotropin (hCG) is given when the largest follicle reaches or exceeds 20-mm mean diameter. The culture media used are modifications of Ham's F-10. Mature oocytes are preincubated for 5 hr before insemination with 50,000 motile sperm per oocyte. Using these protocls, the clinical pregnancy rate is 11.6% per laparoscopy (11/95) and 16.2% per embryo transfer (11/68).     

影响不孕妇女IUI治疗成功率的因素分析

目的:探讨影响IUI治疗成功的各种临床因素。方法:回顾性分析260例接受IUI治疗妇女的促排卵方案、子宫内膜厚度、优势卵泡的数目和直径、IUI的时机、输卵管壶腹部的直径、输卵管伞端距宫角的距离、洗涤后精子的密度和精子的动力、IUI的周期数、AsAb与妊娠结局的关系。结果:IUI治疗结局与子宫内膜的厚度、IUI治疗的时机、输卵管壶腹部的直径、输卵管伞端距宫角的距离、洗涤后精子的密度和动力有关(P<0.01,P<0.05)。与促排卵方案、优势卵泡的数目和直径、抗精子抗体无关(P>0.05)。1-3个周期IUI治疗的妊娠率明显高于3个以上治疗周期的妊娠率(P<0.05)。结论:影响IUI结局的主要因素是:子宫内膜的厚度、IUI治疗的时机、输卵管壶腹部的直径、输卵管伞端距宫角的距离、洗涤后精子的密度和动力。延长IUI治疗的周期数,并不能提高病人的成功率。     

Analysis of factors influencing pregnancy rates in homologous intrauterine insemination

OBJECTIVE: To identify predictors of pregnancy rate (PR) among women undergoing homologous IUI. DESIGN: Cross-sectional analysis of IUI cycles carried out from January 2000 to September 2002. SETTING: Private infertility center in Alicante, Spain. PATIENT(S): Four hundred seventy women undergoing 1,010 cycles of IUI. INTERVENTION(S): Single IUI with ovarian stimulation using hMG. MAIN OUTCOME MEASURE(S): Preovulatory follicles (>15 mm), motile spermatozoa count, type and duration of infertility, female age, insemination timing, and cycle number. RESULT(S): Overall PR per cycle and multiple pregnancy and miscarriage rates were 9.2%, 8.6%, and 11.8%, respectively. Three significant predictors of pregnancy were identified by multiple logistic regression analysis: preovulatory follicles, spermatozoa count, and infertility duration. Interuterine insemination with three follicles almost tripled the PR with respect to only one, odds ratio (OR) = 2.89 (95% confidence interval [CI], 1.54-5.41). Compared with insemination with a motile sperm count >30 x, 20.1-30, 10.1-20, 5.1-10, and < or =5 x10(6), insemination progressively decreased the PR, from 15.3% in the highest category to 3.6% in the lowest (OR lowest/highest = 0.20 [95% CI: 0.09-0.45]), with a statistically significant dose-response trend. Infertility duration > or =3 years was marginally associated with a lower PR, OR = 0.65 (95% CI, 0.40-1.04). Overall, female age was not a significant predictor of pregnancy, and although PR slightly decreased beyond two IUI cycles and when a single IUI was performed 36-40 hours after hCG administration, results were not statistically significant. CONCLUSION(S): Homologous IUI achieves the best results with two or three induced follicles, a high motile spermatozoa count, and infertility duration <3 years, irrespective of female age and fertility history.     

Comparison of fecundability with fresh and frozen semen in therapeutic donor insemination

A series of 401 therapeutic donor insemination (TDI) cycles was analyzed for determining the effect on fecundability of fresh versus cryopreserved semen, the number of inseminations per cycle, recipient age, and the reproductive health of the TDI recipient. We followed a protocol in which inseminations were performed during the periovulatory period determined by urinary luteinizing hormone surge or ultrasound-timed human chorionic gonadotropin injection. The minimum standard for insemination with fresh or cryopreserved semen was a total of 40 X 10(6) grade 3 sperm. We found no decreased fecundability when using frozen semen, rather than freshly ejaculated specimens. The number of inseminations per cycle did not affect the pregnancy outcome after TDI. The reproductive health of the recipient had a significant effect on the pregnancy rate. Fecundability was 15.6% for healthy women, as compared with 6.7% for women with a diagnosis of moderate, severe, or extensive endometriosis, pelvic adhesions, tubal disease, or ovulatory dysfunction. To avoid transmission of human T-lymphotropic virus type III from sperm donor to TDI recipient, we have recently utilized an exclusively frozen TDI program. With well-timed inseminations of 40 X 10(6) grade 3 motile sperm, we have been able to attain pregnancy rates similar to those previously obtained with fresh samples.     

Bovine serum albumin (BSA) can replace patient serum as a protein source in an in vitro fertilization (IVF) program

Alternate protein sources have been suggested to replace the commonly used cord or patient serum for in vitro fertilization (IVF) procedures. During an 11-month period 127 patients treated for in vitro fertilization had elther their serum (N= 71) or bovine serum albumin (BSA;N= 56) used as the protein source in the insemination and growth media. Ham's F-10+0.5% BSA was used for sperm swim-up and insemination media and 1% BSA was used for the growth media. Patient's serum was added to Ham's F-10 culture media at concentrations of 7.5 and 15% for insemination and growth, respectively. Embryo transfer was performed with Ham's F-10 containing 90% maternal serum in both groups. Fertilization rate of 259 oocytes inseminated in medium containing patient's serum did not differ when compared with 200 oocytes inseminated in medium containing BSA. Likewise, rates of obnormal fertilization, cleavage, and pregnancy were similar in both groups. In a second experiment, 148 normally fertilized oocytes were transferred after 24 hr in culture to growth media containing two different concentrations of BSA (0.5 or 1%). Cleavage rates for the two groups were similar and the percentage of embryas developed to 4 cells did not differ signficantly. We conclude that a single concentration of BSA can safely be used to supplement culture media in human IVF with several practical and economical benefits.     

Development of two-cell mouse embryos in protein-free and protein-supplemented media

Two-cell mouse embryos from CFW (Swiss-Webster) mice were cultured to the blastocyst stage in Ham's F10, Whittingham's T6, or human tubal fluid medium. Media were used without any protein supplements or were supplemented with human maternal serum, human fetal cord serum, or human serum albumin. Blastocysts were transferred to modified Eagle's basal medium supplemented with 10% fetal bovine serum for postblastocyst development. Blastocyst and postblastocyst development was depressed among embryos cultured during the preimplantation stage in protein-free Whittingham's T6 and human tubal fluid media compared with embryos cultured in protein-free Ham's F10 medium. This advantage of Ham's F10 disappeared when amino acids and vitamins were added to the other two media. Whittingham's T6 and human tubal fluid supplemented with human serum albumin, human maternal serum, or human fetal cord serum also supported excellent embryo development. When supplermented with protein, Ham's F10 was the poorest of the media in supporting embryo development. Although these results suggest that Ham's F10 is not the best medium for culture of mouse embryos, there is need for caution in extrapolating results from the mouse to the human     

Spermatozoal characteristics from fresh and frozen donor semen and their correlation with fertility outcome after intrauterine insemination.

OBJECTIVE: To determine if conventional sperm parameters, specific characteristics of sperm motion determined by computer-aided semen analysis (CASA), sperm penetration assay (SPA), and/or spontaneous acrosome reaction assay could best predict fertility outcome after intrauterine insemination (IUI) from frozen donor sperm. DESIGN: A retrospective analysis of 2,245 cycles of therapeutic donor IUIs were initially studied; 1,147 cycles that met selection criteria were used in this report. SETTING: A university-based assisted reproductive technology center. PATIENTS, PARTICIPANTS: All IUIs were performed on women with documented patency of at least one fallopian tube, ovulatory cycles, and who did not receive human menopausal gonadotropins. Sperm donors had to be used for at least four different recipients (mean of 15) and at least 14 different cycles of insemination (mean of 41). INTERVENTIONS: None. MAIN OUTCOME MEASURE: Pregnancy. RESULTS: Statistical comparisons were made between donors of different relative fertility by using the Mann-Whitney test, Spearman's rank correlation, and multiple regression analysis. These analyses demonstrated that the most significant predictors of the fertility of frozen-thawed donor sperm were curvilinear velocity, straight line velocity, and the total number of motile sperm inseminated. The number of sperm with spontaneous acrosome reactions negatively correlated with fertility outcome, and SPA provided no predictive value. CONCLUSIONS: Our study supports the hypothesis that the study of sperm motion characteristics using CASA after thawing and washing of cryopreserved sperm is a better predictor of fertile outcome after IUI than analysis of fresh semen.     

Improved pregnancy rates and outcome with gamete intrafallopian transfer when follicular fluid is used as a sperm capacitation and gamete transfer medium

Follicular fluid (FF) is a dynamic medium rich in steroids, polypeptide hormones, and growth factors. Preovulatory FF can stimulate spermatozoal acrosome reaction. Moreover short preincubation of washed sperm with FF improves sperm performance in the hamster egg penetration assay. In the current study, FF was used to capacitate sperm and as a transfer medium in 131 gamete intrafallopian transfer (GIFT) procedures. Ham's F-10 medium (GIBCO, Grand Island, NY) with 50% maternal serum was used in another 29 GIFT procedures. In the Ham's F-10 group, 29 GIFT procedures were performed in 25 patients with a pregnancy rate of 21% per laparoscopy and 24% per patient. In the FF group, 131 GIFT procedures were performed in 100 patients with a pregnancy rate of 50% per laparoscopy and 66% per patient, with a 59% ongoing pregnancy rate per patient. The most significant improvement was in the male factor group with a pregnancy rate of 0% in the Ham's F-10 group and 44% per procedure in the FF group. We conclude that the use of FF for sperm capacitation and as a gamete transfer medium significantly improves GIFT pregnancy rates and outcome.     

Intrauterine insemination outperforms intracervical insemination in a randomized, controlled study with frozen, donor semen.

OBJECTIVE: To assess the efficacy of intrauterine insemination (IUI) in a donor insemination program. DESIGN: Prospective randomized clinical trial. SETTING: Donor insemination program. PATIENTS, PARTICIPANTS: Women undergoing insemination were randomly assigned to receive either IUI or intracervical insemination for a maximum of six cycles. INTERVENTIONS: None. MAIN OUTCOME MEASURE(S): Cycle fecundity rates between the two routes were compared. RESULTS: The monthly fecundity rate for intracervical insemination was 5.1% compared with 23% by IUI. By life table analysis, pregnancy rates for IUI were significantly higher than intracervical insemination (P = 0.02). CONCLUSIONS: Intrauterine insemination with quarantined donor sperm is superior to intracervical insemination.     

A simple method for fallopian tube sperm perfusion using a blocking device in the treatment of unexplained infertility

Objective: To evaluate the efficacy of fallopian sperm perfusion (FSP) using a new method similar to the FAST system® in comparison with standard intrauterine insemination (IUI) in patients with unexplained infertility.

Design: Prospective, randomized, controlled study.

Setting: Assisted conception service in a University Hospital.

Patient(s): Women with unexplained infertility undergoing controlled ovarian hyperstimulation (COH).

Intervention(s): After hCG administration, patients were randomized to either standard IUI or FSP. The women received the same treatment in the first and all subsequent cycles. A maximum of three cycles was performed. Intrauterine insemination was performed using a standard method, and fallopian sperm perfusion was performed using a commercial device for hysterosalpingography and tubal hydropertubation.

Main Outcome Measure(s): Clinical and ongoing pregnancy rates.

Result(s): A total of 132 cycles was completed: 66 IUI cycles and 66 FSP cycles. In the IUI group, there were 5 ongoing pregnancies, giving a pregnancy rate of 7.6 per cycle and 15.6% per patient; in the FSP group, 14 ongoing pregnancies occurred, giving a pregnancy rate of 21.2% per cycle and 42.4% per patient. The prevalence of multiple pregnancies, miscarriages and ectopic pregnancies was similar in the two insemination groups. Fallopian sperm perfusion was easy to perform, and no case of sperm reflux was observed. The procedure was well tolerated and no complications were observed. The costs were comparable with standard IUI.

Conclusion(s): In the treatment of couples with unexplained infertility, the method for fallopian sperm perfusion described yields higher pregnancy rates than IUI, with no significant increase in costs or complications. However, these results need to be confirmed in larger studies before replacing IUI with FSP as standard practice.     

A prospective randomized study of pregnancy rates following intrauterine and intracervical insemination using frozen donor sperm

Cryopreserved sperm have lowered fertility when compared with fresh sperm in artificial insemination by donor programs. The purpose of this study was to compare pregnancy rates following intrauterine insemination (IUI) and intracervical insemination (ICI) with cryopreserved sperm in a prospective trial using the patient as her own control. A total of 154 patients were randomized into alternating treatment cycles and underwent 238 cycles of IUI and 229 cycles of ICI. The pregnancy rate per treatment cycle was 9.7% following IUI and 3.9% following ICI. Treatment outcome was influenced by patient age, ovulatory status, and endometriosis. Pregnancy success correlated well with the post-thaw survival of sperm and the number of motile cells inseminated. In spite of having normal semen parameters, some donors were found to have markedly reduced sperm fecundity. We conclude that IUI with cryopreserved sperm can be an effective treatment for couples with infertility, genetic indications, or other reasons.