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PubMed2001

Induction of follicular growth and ovulation with urinary and recombinant gonadotrophins.

Gonadotrophins have been used since the 1930s. By the 1990s, a pure biological extract of FSH was obtained. The increase in the demand for new assisted reproduction techniques led to the creation of recombinant FSH, 99% pure and highly consistent between batches, with no LH activity, and a high specific activity. In gonadotrophin protocols, follicular growth can be monitored with regular ultrasounds and/or hormonal testing, mainly oestradiol in the follicular phase. In assisted reproduction the adoption of ovulation induction strategies with GnRH analogues, control the endogenous secretion of LH, avoiding cancellations due to an early LH surge, and premature ovulation. Depending on the moment when pituitary suppression starts, there are short, ultra-short and long protocols The use of long protocols not only increases pregnancies and live births, it also allows an easier programming and simplifies IVF treatments. Ovarian stimulation entails some risks such as ovarian hyperstimulation syndrome and multiple pregnancies that can be avoided in many cases with an accurate prediction and an active prevention.

Journal Article↗

PubMed1998

Artificial insemination in Callithrix jacchus using fresh or cryopreserved sperm.

Assisted reproductive techniques are needed urgently to facilitate the captive breeding of many New World primate species which are endangered in the wild and to assist the effective genetic management of small colonies. A protocol was devised for artificial insemination in the common marmoset, Callithrix jacchus, using ejaculated sperm obtained by vaginal washing after copulation. A double insemination protocol was employed, with the first insemination taking place the day before ovulation was expected to occur and the second 48 h later. All six females inseminated with fresh ejaculated sperm became pregnant, delivering a total of 16 offspring at term. The gestation lengths and litter sizes were not statistically different from those observed in pregnancies following natural mating. The insemination protocol was adapted for use with cryopreserved ejaculated sperm by including an additional insemination on the day of expected ovulation, to take into account differences in the capacitation time of frozen-thawed sperm compared to fresh sperm. Three out of six females inseminated according to this triple insemination schedule, conceived, although one female subsequently resorbed twin foetuses approximately 100 days later. The remaining two pregnant females delivered four babies at term, one singleton and one set of triplets. In the final group, six females were inseminated with low doses of cryopreserved epididymal sperm using the same triple insemination protocol used for frozen-thawed ejaculated sperm. One female conceived, delivering triplets.

Animals↗

PubMed1997

A new method for the purification of human motile spermatozoa applying density-gradient centrifugation: polysucrose media compared to Percoll media.

PURPOSE: A newly developed method for the isolation of human motile spermatozoa using density-gradient centrifugation was compared with the traditionally used Percoll technique. METHOD: Sperm samples were divided into two equal aliquots, which were purified with either the traditionally performed Percoll technique or a new alternative based on polysucrose/Optiprep media. For each sample the isolation was performed during the same run of the centrifuge. RESULTS: The average recovery of progressively motile spermatozoa with the polysucrose/Optiprep method was significantly higher (48 +/- 7%) than with the Percoll method (38 +/- 6%) (n = 18). The average percentage of motile spermatozoa and the motility score were similar in the purified preparations. CONCLUSIONS: The new polysucrose/Optiprep-based density-gradient centrifugation technique for the isolation of motile human spermatozoa is as good as the traditionally used Percoll method and may replace it in connection with assisted reproduction techniques.

Cell Separation↗

PubMed1991

Pregnancy from cryopreserved embryos: an important new aspect of assisted reproduction.

Assisted Reproduction techniques have become well-established methods of treating infertility in the decade since their inception. The gradually improving efficacy of these procedures has made the risks of multiple pregnancy a serious concern. Cryopreservation of "extra" embryos has been developed to counter this problem, but has led to new difficulties. Assisted Reproduction in Kentucky has kept pace with progress worldwide; the first Kentucky live birth from transfer of cryopreserved embryos is reported.

Adult↗

PubMed1994

Effects of pentoxifylline and progesterone on human sperm capacitation and acrosome reaction.

This study was designed to test the effects of pentoxifylline and progesterone upon capacitation of fresh human spermatozoa. Capacitation and acrosomal integrity were assessed using the fluorescent probe chlortetracycline on spermatozoa co-stained with a supravital fluorescent dye, Hoechst 33258. Hyperactivated motility was measured using computer-assisted movement analysis. After exposure to pentoxifylline (1 mg/ml; 30 min), the fluorescent 'B' pattern, characteristic of capacitated, acrosome-intact cells, increased significantly (P < 0.01), though no increase in 'AR' pattern, characteristic of acrosome-reacted cells, was detected. There was a significant increase in hyperactive motility (P < 0.001). Exposure to progesterone (1 microgram/ml; 60 min) resulted in a significant increase in 'B' pattern (P < 0.05) and 'AR' pattern (P < 0.005), though no effect on the expression of hyperactivation was detected. No effect upon hyperactivation was detected on exposure of fresh or cryopreserved spermatozoa to a physiological range of progesterone concentrations (0.1-1000 ng/ml). Sequential exposure to pentoxifylline then progesterone resulted in a significant increase in 'B' pattern, acrosome loss and hyperactivation. Sperm viability was not affected in any treatment group. These observations suggest that pentoxifylline and progesterone affect capacitation through independent mechanisms. Stimulation of both capacitation and acrosome reaction resulted from sequential exposure to pentoxifylline and progesterone. This may have implications for sperm handling for assisted reproductive techniques.

Acrosome↗

PubMed2006

[Cancer and fecundity].

The overall increase in relative survival rates of adult and childhood cancer patients is related to improved chemotherapy and radiotherapy treatment protocols. However, this success is tempered by gonadotoxic effects and uterine damage. The reduced fertility and possible premature menopause can have a profound impact on patients self-esteem and quality of life. Consequently, preservation of fertility or treatments to protect the ovarian function, become a more relevant issue. Advances in assisted reproduction techniques have focused on the following: embryo cryopreservation which is restricted by time and constrains such as, sexual maturity and the presence of a stable partner. Cryopreservation of immature ovarian tissue, the most promising option, with restoration of natural fertility following auto-transplantation or maturation of oocytes in vitro in combination with assisted reproduction.

Cryopreservation↗

PubMed2005

Sperm washing techniques address the fertility needs of HIV-seropositive men: a clinical review.

It is estimated that nearly 40 million people worldwide are infected with HIV. HIV/AIDS prevalence among young people is high, with youth under age 25 years accounting for approximately half of all new infections. Nearly 1 million Americans are HIV-seropositive. Today, HIV-seropositive individuals live active and productive lives despite their illness, largely a result of prescribed highly active antiretroviral therapy (HAART). Most individuals are of reproductive age, and many desire to have children. Various assisted reproductive techniques have been used to reduce or eliminate infectious elements known to be present in semen so that isolated spermatozoa can be safely inseminated or used for intracytoplasmic sperm injection into oocytes. Presently, several European centres and a few US groups offer assistance to HIV seropositive men and their seronegative partners by performing either intrauterine insemination (IUI) or IVF. Since 1987, more than 3600 published attempts have been reported in which processed spermatozoa from HIV-seropositive men were used to establish pregnancy in HIV-seronegative women. Although the data remain observational, sperm washing techniques appear to be relatively safe and effective, offering HIV-serodiscordant couples an opportunity to have children.

Fertilization in Vitro↗

PubMed2004

The Developing Country Reactions to Biomedical Techniques and Plant Biotechnology: The Tunisian Experience.

In the present study we present the conditions offered to biotechnology development in Tunisia and we compare three main biotechnology applications which raise ethical and health problems: organ transplant, assisted reproductive techniques, and genetically modified organisms. We try to identify factors that have allowed success of the first two applications and failure of the latter. Conditions offered to biotechnology in other African countries are also discussed.

Journal Article↗

PubMed1998

Pregnancy achieved by intracytoplasmic sperm injection using cryopreserved vasal-epididymal sperm from a man with spinal cord injury.

Anejaculation and poor semen quality are two major causes of infertility in men with spinal cord injury (SCI). The poor motility of retrieved sperm usually has low fertilization potential and is thought to be unfavorable for cryopreservation. This report describes a pregnancy after intracytoplasmic sperm injection (ICSI) with cryopreserved vasal-epididymal sperm from a man with SCI and anejaculation. An attempt was made to obtain sperm through electroejaculation, but no motile sperm were found in two trials. Therefore, the subject underwent vasal aspiration. The retrieved sperm had a concentration of 26 x 10(6)/mL and a motility of 3%. ICSI was considered to be the best choice for the couple, but the wife did not become pregnant in the first cycle of treatment. A successful pregnancy was achieved by ICSI in the second cycle using frozen-thawed sperm, supernumerary in the previous cycle, with a density of 5 x 10(6)/mL and 1% motility. A set of healthy twins, one boy and one girl, were delivered via cesarean section at 36 weeks of gestation. Complementary to other assisted reproductive techniques, ICSI may provide men with SCI a greater opportunity to father children. The supernumerary sperm, regardless of quality, should be cryopreserved to avoid the necessity and risk of repeated assisted ejaculations and aspirations of the genital tract.

Adult↗

PubMed1993

Electroejaculation.

Ejaculatory dysfunction is a relatively uncommon cause of male infertility. In men with total anejaculation and in some men with psychogenic anejaculation, electroejaculation is a very useful treatment. By following the guidelines provided, electroejaculation can be performed reproducibly and safely with a good success rate. Deviations from the safety techniques detailed may result in unsafe procedures. The risks involved with performing electroejaculation recklessly can be significant, including rectal injury and cerebrovascular accident from hypertension. Pregnancy rates from electroejaculation coupled with assisted reproductive techniques are not optimal, and much work continues at our center to determine why this is the case.

Ejaculation↗

PubMed1992

[What results can be expected from in vitro fertilization and other technics of assisted fertilization?].

Results of assisted reproductive techniques (IV, GIFT and ZIFT) have improved progressively during the last years but they are still not satisfactory specially when they are analyzed in terms of cost efficacy. During 1990, in the world literature, the following successful results were reported: pregnancy rates of 20% and delivery rates of 15% for IVF-ET, 29% and 22% for GIFT and 21% and 16% for ZIFT. In spite of the small number of cases reported in Chile, results are similar to those of USA and Europe. Results of assisted fertilization techniques have improved with the use of Gn-RH agonists and are worse in patients older than 35 years of age than in younger ones. Pregnancies rates are higher when the number of transferred ovocytes increases although transfer of more than 3 or 4 ovocytes or embryos is contraindicated because of the risks of multiple pregnancies. When more than 3 or 4 embryos are obtained those not transferred are usually cryopreserved. IVF-ET is the first choice of treatment in cases of infertility due to severe tubal damage and/or extensive and dense pelvic adhesions. In cases of infertility with normal tubes, intrauterine inseminations in 3 to 4 controlled hyperstimulated cycles are recommended before indicating an assisted fertilization procedure. The most risky complication of assisted reproduction, besides multiple pregnancy, is severe ovarian hyperstimulation syndrome (SOHS). This syndrome is prevented by not injecting HCG or by not transferring ovocytes or embryos in cases of high estradiol levels after HMG and/or FSH administration.

Adult↗

PubMed1995

Improved techniques for first-trimester fetal reduction using simultaneous abdominal and transvaginal ultrasonographic guidance. Preliminary report.

Simultaneous transabdominal injection of potassium chloride (KCl) assisted by transvaginal ultrasonographic guidance was evaluated in 5 patients who conceived due to various assisted reproductive techniques and ovarian stimulation. All 5 underwent first-trimester fetal reduction for multiplicity of high-order pregnancies. Four sets of triplets were reduced to twin, and 1 twin pregnancy was reduced to a singleton. Less than 2 ml of 2 mEq/ml KCl per fetus was needed for each case. No fetal or maternal complications nor technical failure attributable to the procedure were recorded. All pregnancies were delivered > or = 35 weeks of gestation. Transabdominal fetal reduction using simultaneous transvaginal ultrasonographic guidance for selective feticide combines the benefits of both the transabdominal and transvaginal approaches, mainly in selected cases in which some experts prefer the abdominal route, but find it technically difficult to practice. Further experience is needed to confirm our encouraging preliminary results before drawing any definitive conclusions.

Abdomen↗

PubMed1993

Ejaculatory failure.

Although anejaculation is a relatively uncommon occurrence in the general population, over 12,000 new cases are reported annually. Anejaculation may result from spinal cord injury (SCI), retroperitoneal lymph node dissection (RPLND), diabetes mellitis, transverse myelitis, multiple sclerosis, or psychogenic disorders. At least 30% of men with this problem are or will be married and many will seek help to remedy their infertile state. The evolution of technique and instrumentation over the least 30 years has made electroejaculation an accessible and acceptable form of therapy. Recent successes in inducing ejaculation by means of rectal probe electrostimulation or vibratory stimulation combined with assisted reproductive techniques, such as zygote intrafallopian transfer (ZIFT), gamete intrafallopian transfer (GIFT), and in vitro fertilization (IVF), have provided these men a means of producing their own biologic offspring.

Combined Modality Therapy↗

PubMed2006

Comparison of fallopian tube sperm perfusion and intrauterine tuboperitoneal insemination: a prospective randomized study.

OBJECTIVE: To evaluate the results of two different methods of insemination: fallopian tube sperm perfusion (FSP) with 4 mL of inseminate and intrauterine tuboperitoneal insemination (IUTPI) with 10 mL of inseminate. DESIGN: Prospective randomized clinical study. SETTING: Private infertility center. PATIENT(S): Two hundred seventy-six couples, undergoing 403 cycles, with unexplained infertility, mild or moderate male infertility, or mild or moderate endometriosis. INTERVENTION(S): Patients were assigned randomly to either FSP (group A, n = 138) or IUTPI (group B, n = 138) treatment. Both groups followed the same mild ovarian stimulation protocol. MAIN OUTCOME MEASURE(S): Ninety-five overall pregnancies: 35 in group A (FSP) and 60 in group B (IUTPI). RESULT(S): The pregnancy rate per cycle (presence of gestational sac with heart beats) was 17.6% in group A (n = 199) and 29.4% in group B (n = 204). These differences were statistically significant (P < .007). The odds ratio of getting pregnant, per patient, in group B was 2.26 (95% confidence interval 1.36-3.77) compared with group A. CONCLUSION(S): The results of this study indicate that IUTPI may prove to be a useful technique in the treatment of unexplained infertility, mild or moderate male infertility, and mild or moderate endometriosis. Three attempts of IUTPI may be beneficial before moving on to more invasive and expensive methods of assisted reproduction techniques.

Adult↗

PubMed1990

Co-cultures: their relevance to assisted reproduction.

Assisted reproductive techniques have contributed significantly to alleviating subfertility in the childless couple. However, the take-home baby rates have been very low throughout the world. One of the contributory causes has been the reduced viability of replaced embryos brought about by suboptimal in vitro conditions. The culture of human embryos in the presence of passaged human tubal ampullary monolayers (co-cultures) is an attractive approach to improving the viability of embryos for assisted reproduction. Seventy per cent of blastocysts can be produced in human ampullary co-culture as compared to 33% in standard culture media. This paper discusses the various roles of human co-cultures in assisted reproduction and provides an opinion as to how the transfer of blastocysts produced via the co-culture system could enhance pregnancy rates. Particular emphasis is placed on human oviductal characteristics, the various co-culture systems, screening of co-cultures for microbes, freezing of ampullary cells, growth factors and embryonic blocks, specificity of co-cultures, sperm hyperactivation in co-culture and pregnancy rates. The first patient on a clinical trial who had four of her oocytes fertilized and grown in human ampullary co-culture and then replaced into her uterus became pregnant. The co-culture system may have tremendous potential in supporting human embryonic growth via embryotrophic factors.

Animals↗

PubMed2001

Preoperative semen analysis as a predictor of seminal improvement following varicocelectomy.

OBJECTIVE: To determine the predictive role of preoperative semen analysis on both seminal improvement and pregnancy rates following varicocelectomy. DESIGN: Retrospective data analysis. SETTING: Two academic medical center infertility clinics. PATIENT(S): One hundred ten consecutive patients who underwent varicocelectomies. Seminal improvement data were available for 84 patients, and pregnancy data were available for 58 patients. INTERVENTION(S): Stratification of patients based on preoperative total motile sperm count (TM). Varicocelectomy was performed on all patients. MAIN OUTCOME MEASURE(S): TMs, pregnancy rates, and conception techniques following varicocelectomy of each preoperative group. RESULT(S): Men with mild to moderate oligoasthenospermia (TM >5 million) had significantly better seminal improvement following varicocelectomy. While preoperative stratification showed no difference in pregnancy rates (when assisted reproductive techniques were included), men who achieved a postoperative TM >20 million were more likely to achieve conception by less invasive techniques (natural and intrauterine insemination vs. in vitro fertilization [IVF]). CONCLUSION(S): Varicocelectomy may be the most cost-effective initial intervention in males with TM >5 million. Patients with TM <5 million and concomitant female factor infertility may be better initial candidates for IVF.

Adult↗

PubMed2005

Oxidative stress and its implications in female infertility - a clinician's perspective.

Reactive oxygen species (ROS) have a role in the modulation of gamete quality and gamete interaction. Generation of ROS is inherent in spermatozoa and contaminating leukocytes. ROS influence spermatozoa, oocytes, embryos and their environment. Oxidative stress (OS) mediates peroxidative damage to the sperm membrane and induces nuclear DNA damage. ROS can modulate the fertilizing capabilities of the spermatozoa. There is extensive literature on OS and its role in male infertility and sperm DNA damage and its effects on assisted reproductive techniques. Evidence is accumulating on the role of ROS in female reproduction. Many animal and human studies have elucidated a role for ROS in oocyte development, maturation, follicular atresia, corpus luteum function and luteolysis. OS-mediated precipitation of pathologies in the female reproductive tract is similar to those involved in male infertility. OS influences the oocyte and embryo quality and thus the fertilization rates. ROS appears to play a significant role in the modulation of gamete interaction and also for successful fertilization to take place. ROS in culture media may impact post-fertilization development, i.e. cleavage rate, blastocyst yield and quality (indicators of assisted reproduction outcomes). OS is reported to affect both natural and assisted fertility. Antioxidant strategies should be able to intercept both extracellular and intracellular ROS. This review discusses the sources of ROS in media used in IVF-embryo transfer and strategies to overcome OS in oocyte in-vitro maturation, in-vitro culture and sperm preparation techniques.

Antioxidants↗

PubMed1992

Current status of in vitro fertilization and the new reproductive technologies.

The rapid pace of development and refinement of in vitro fertilization and ancillary assisted reproductive techniques warrants periodic review to keep the reader abreast of the growth in this clinical and scientific arena. Although not exhaustive, this review targets key areas of special interest and significance, including oocyte and embryo micromanipulation for assisted fertilization and genetic analysis, advances in the approach to ovarian stimulation, and examination of the scientific model afforded by ovum donation.

Female↗