[What strategy to improve the recruitment of gamete donors?]. / Quelle stratégie pour améliorer le recrutement des donneurs de gamètes?

To a high demand for donated gametes, sperm and oocytes, centers often face difficulties in recruiting enough donors. To increase the number of donors are needed financial and human resources to accommodate them in good conditions, information and a strong promotion to motivate approaches donations, diversify recruitment methods. In France, the gametes donation comes from couples with children and is free. The opening of the gift for men and women without children and remuneration of these gifts are discussed.

[Anonymity and gamete donation]. / Anonymat du don de gamètes.

In France the gamete donation is based on the major principles: anonymity, no payment, solidarity and this mode of procreation can be used only if a medical indication is present in recipient couples. In prerequisite and during the revision of the law of bioethics, a wide debate took place about the anonymity of gamete donation. The objectives of this article is to review the concept of the anonymity and its links with the questions of the origin, the secret of the modalities of the conception and the mourning of the fertility, children, donors and the recipient couples waiting for gamete donation. The international situation is also addressed. The contribution of the CECOS, the centers which practice the sperm and the egg donations is highlighted. The anonymity cannot be discussed without addressing all these links and the complexity of this particular mode of conception. To date, the French society has maintained the anonymity in the new law of bioethics.

[Secrecy and anonymity in gametes donation]. / Secret et anonymat du don de gamètes.

In France, gamete donation is based on some fundamental principles such as anonymity, gratuity, and solidarity. In Europe a trend appeared for a removal of anonymity. This paper highlights the reasons for this removal of anonymity, and consequences for the donor's offspring, the gamete donor, the infertile couples. In particular, an examination of practice shows that removing the anonymity of gamete donation favors maintaining the secrecy of the circumstances of the conception, resulting in inefficiency of the law.

[Oocyte donation as in France]. / Pour un don d'ovocytes à la française.

Oocyte donation, initially proposed in agonadal women, saw indications expand to ovarian deficiencies and failures of in vitro fertilization (IVF), resulting in a significant increasing demand. The recruitment of oocyte donors is a critical issue for all countries that have allowed this practice. The French legislation, with the laws of bioethics, is clearly the most restrictive of European countries, imposing an absolute free gift from mother. The different solutions in the neighboring countries are analysed and in particular the interpretations made in respect of gratuity and compensation. Motivating donors (spontaneous, relational, or by reciprocity), but also motivating the medical teams can organize a program of oocyte donation in France. The authors present their results of three years experience, demonstrating that this system is possible in the current legislative framework.

[Single-embryo transfer: Rennes' Hospital experience]. / Transfert monoembryonnaire : expérience du CHU de Rennes.

In 2005-2006, 905 punctures of oocytes were realised in the Assisted Reproductive Technology Centre of Rennes' Hospital in France, the source of 173 pregnancies after fresh embryos transfers and 185 pregnancies after frozen embryos transfers. The single-embryo transfer (SET) was proposed in all patients aged less than 38 years with at least two embryos type I or II on the first two cycles. Sixty-three percent of patients (n=293 cycles) chose the SET with 21.5% pregnancies through fresh embryo transfer. The SET with frozen embryos has been completed on 708 cycles with 16.8% of pregnancies per transfer. The cumulative rate of pregnancies by puncture is 39% and the overall risk of multiple pregnancies has dropped to 12%. The SET associated with an effective embryo cryopreservation therefore allows to reduce the risk of multiple births while maintaining a satisfactory pregnancy cumulative rate.

[Intrauterine insemination: against systematic ovarian stimulation]. / Insémination intra-utérine: contre une stimulation systématique de l'ovulation.

Intrauterine insemination is a technique proposed of first intention in many situations of non-tubal infertilities. It is very frequently associated with a stimulation of ovulation, with the objective of precisely synchronizing the intra uterine stimulation and ovulation, and an increase in follicular recruitment. If intrauterine stimulation is often a determining factor in obtaining pregnancy, stimulation of ovulation does not always bring about an obvious benefit and its interest must be discussed according to the indications, knowing on the other hand that it is determining in the occurring of multiple pregnancies.

Geographic variations in probability of pregnancy in four cities of France.

BACKGROUND: Several studies have described geographic variations in human fecundability, but this phenomenon has almost exclusively been studied at an international level rather than within a given country. Our aim was to describe geographic variations in fecundability, the monthly probability of pregnancy, between four cities of France. METHODS: We conducted a cross-sectional study in four French maternity units from Toulouse, Rennes, Lyons and Paris, among partners of pregnant women. Women were asked about the time to pregnancy (TTP) of their current pregnancy. TTP was analysed with a discrete Cox model allowing to estimate fecundability ratios (FR). RESULTS: Time to pregnancy was defined for 894 couples. There was no strong evidence of heterogeneity in fecundability between the four compared cities (p=0.05 without adjustment and p=0.25 after adjustment for behavioural and medical factors). The highest fecundability was observed in Rennes and the lowest in Toulouse (fecundability ratio (FR)=1.28, 95% CI: 1.01-1.63). Differences in fecundability were smaller between the other cities. CONCLUSION: We highlighted a possibly slightly higher fecundability in Rennes compared to Toulouse. Possible explanations for this finding are discussed. We note that the finding is consistent with previous observations indicating a higher sperm concentration among semen donors in Rennes than in Toulouse.

[Artificial insemination with donor frozen sperm]. / Insémination artificielle avec sperme congelé de donneur.

The cryopreservation of human sperm decreases its fecondity, by reduction in the number of motile sperm. However the fertilizing ability of the spermatozoa is not faded. The use of frozen sperm requires an optimization of the methods of artificial insemination to hope to approach the results obtained with fresh sperm. Intrauterine insemination with sperm selection meets this aim.

Selection bias in semen studies due to self-selection of volunteers.

BACKGROUND: Reports of a secular decrease in semen quality remain controversial, particularly due to the possibility of selection bias. We aimed to describe the potential bias due to self-selection of volunteers in semen studies involving fecund men. METHODS: Using data from the French multicentre study REPRHOM, we compared the characteristics of the partners of pregnant women for three levels of participation: completion of a refusal questionnaire (n = 698), agreement to complete the study questionnaires only (n = 676) and agreement to complete the study questionnaires and give a semen sample (n = 331, 13% of the subjects approached). RESULTS: Poorly educated men refused more often to participate than highly educated men. Semen providers were more likely to have experienced unfavourable pregnancy outcomes (odds ratio 1.68, 95% confidence interval 1.14-2.49) compared with participants completing the questionnaires only. Time to pregnancy was similar for all participants. CONCLUSIONS: This study demonstrates the existence of selection bias in semen studies associated with fertility and socio-demographic characteristics of men. The results of semen analysis for this population sample cannot be extrapolated to the whole population from which the volunteers originate. More information is required on who participates, and participation rates should be reported in semen studies to make it possible to interpret the results correctly.

[Is the limitation to 6 cycles of insemination with donor sperm justified?]. / La limitation à six cycles d'inséminations en AMP avec don de sperme est-elle justifiée?

OBJECTIVE: To evaluate the effect of the limitation at 6 cycles of artificial insemination in a program of sperm donation. STUDY DESIGN: 266 couples included in a program of sperm donation underwent 1,354 cycles including 532 intracervical insemination (ICI), 678 intrauterine insemination (IUI) and 133 in vitro fertilization (IVF). RESULTS: The birth rate by cycle was 10.8% in ICI, 18% in IUI, 21.9% in IVF. The risk of multiple pregnancies was 0% in ICI, 13% in IUI, 33% in IVF. The authors feign then two strategies, the first one with 6 ICI followed by 6 IUI, and the other one with 6 IUI alone. The birth rate, the risk of multiple pregnancies and the cost of these two strategies is discussed. CONCLUSION: The restriction to 6 IA Cycles in a donor semen program does not change the birth rate, but increases the multiple pregnancy rate and the cost of the treatment of these patients.

Molecular screening of the CFTR gene in men with anomalies of the vas deferens: identification of three novel mutations.

Many studies have shown that congenital absence of the vas deferens (CAVD) is a genital cystic fibrosis transmembrane conductance regulator (CFTR)-mediated phenotype, with a broad spectrum of abnormalities causing male infertility. The genotype of these patients includes mutations in the CFTR gene, e.g. DeltaDeltaF508, R117H and the T5 allele; all of which are commonly found in CAVD. In this study we have screened the entirety of CFTR gene in 47 males with anomalies of the vas deferens: 37 cases of congenital bilateral absence of the vas deferens, three cases of congenital unilateral absence of the vas deferens and seven cases of obstructive azoospermia with hypoplastic vas deferens. Among the 94 chromosomes studied, 65 mutations, of which three are novel (2789+2insA, L1227S, 4428insGA), were identified. The majority of patients (63.8%) had two detectable CFTR gene mutations. Furthermore, high frequencies of the DeltaDeltaF508 mutation (44.7%), the T5 allele (36.2%) and R117H mutation (19.1%) were observed.

[Microsurgical epididymal sperm aspiration (MESA), testicular biopsy and intracytoplasmic sperm injection (ICSI) in the treatment of male infertility]. / Aspiration microchirurgicale des spermatozoïdes épididymaires (MESA), biopsie testiculaire et injection intra-cytoplasmique du spermatozoïde (ICSI) dans le traitement de l'infertilité masculine.

OBJECTIVE: To assess if ICSI using epididymal or testicular spermatozoa is effective in the treatment of couples with male factor infertility. MATERIAL AND METHODS: 56 couples suffering from male infertility underwent a total of 88 treatment cycles of ICSI, in combination with microsurgical epididymal sperm aspiration (MESA): 62 cycles, or testicular sperm retrieval: 26 cycles. RESULTS: Overall, fertilization rate was 63% per injected metaphase II oocyte. In MESA group, fertilization rate was 64% compared to 61% embryos in testicular sperm aspiration. The overall pregnancy rate was 26% per started ICSI cycle. Pregnancy rates were also similar in both group: 24% for MESA and 32% for testicular sperm extraction. 9 pregnancy (41%) were obtained using cryopreserved epididymal sperm (6 cases) or testicular sperm (3 cases). CONCLUSION: ICSI combined with epididymal or testicular sperm achieved a high fertilization and pregnancy rate. It constitutes an efficient alternative in the treatment of men suffering of testicular failure or azoospermia not amenable to surgical reconstruction.

Influence of seminal plasma on cryopreservation of human spermatozoa in a biological material-free medium: study of normal and low-quality semen.

The objective was to evaluate the efficiency of a biological material-free medium and the role of seminal plasma (SP) in the cryopreservation of human spermatozoa. Normal semen samples and low-quality semen samples were used for this study. After centrifugation of 300 microL fractions of whole semen, pellets were resuspended either in autologous SP or in a chemically defined medium (BM) supplemented or not with 3% bovine serum albumin (BSA); after 15 min at 37 degrees C, the samples were diluted (V/V) with cryoprotective medium (30 mM NaCl; 22 mM sodium citrate, 19.4 mM fructose; 80 mM glutamine; 14%, V/V, glycerol) and maintained for 15 min at room temperature before freezing. Assessment of viability and motility was performed using fresh semen (T0), after centrifugation and resuspension prior to adding the cryoprotectant (T15), after adding the cryoprotectant (T30) and after freezing and thawing (Tpost). In all three resuspending media used, sperm viability and motility (forward and total) decreased (p < 0.05) during both the equilibration period especially before addition of the cryoprotective medium (between T0 and T15) and during the freeze-thaw process comparison between T30 and Tpost. The recovery of viable and motile spermatozoa (post-thaw values/values of fresh samples) was higher (p < 0.05) in normal semen than in low-quality semen. In both groups, the recovery was slightly, but significantly, higher with SP than with BM and the presence of BSA has no beneficial effect. To conclude, these data suggest that SP may reduce the deleterious effects of cryopreservation. Nevertheless cryopreservation of spermatozoa in a medium containing neither SP nor biological substances could offer an acceptable cryoprotection of spermatozoa to be used in assisted fertilization procedures, especially for intracytoplasmic sperm injection.

Influence of oxygen tension on function of isolated spermatozoa from ejaculates of oligozoospermic patients and normozoospermic fertile donors.

Oxygen radical generation is known to be detrimental to sperm function. An example of a reactive oxygen species-associated male pathology is oligozoospermia in which fertilization and pregnancy rates are low in in-vitro fertilization (IVF) programmes. As the extent of the modifications induced by reactive oxygen species (ROS) depends on several factors, notably from oxygen tension in the incubation medium, the aim of this study was to examine the influence of a low (5%) rather than atmospheric (20%) oxygen tension in the incubator gas phase on the function of Percoll-selected spermatozoa from ejaculates of oligozoospermic patients and normozoospermic fertile donors. After incubation for several hours in a gas phase of either 5% CO2/90% N2/5% O2 or 5% CO2/95% air (20% O2), none of the parameters investigated, e.g. movement characteristics, potential of spermatozoa to acquire hyperactivated motility, to undergo the acrosome reaction when challenged with a calcium ionophore and to fuse with zona-free hamster oocytes, was significantly different between the two oxygen tensions in fertile donors. In contrast, among oligozoospermic patients, the motility parameters, the percentage of hyperactivated motility and of induced-acrosome reaction were significantly improved under a gas phase of 5% O2 compared with those observed under an atmosphere of 20% O2 (P < 0.05). Exposure to 5% rather than 20% oxygen tension also induced a significant increase in the percentage of penetration of zona-free hamster eggs after capacitation for 17 h, but no difference was found in the mean number of bound spermatozoa per oocyte. After incubation for 24 h, a significantly higher survival rate was observed under 5% compared with 20% oxygen tension. These results show that the use of a low oxygen tension rather than air might improve spermatozoan competence of oligozoospermic patients during IVF programmes.

Multicentre approaches to donor insemination in the French CECOS Federation: nationwide evaluation, donor matching, screening for genetic diseases and consanguinity. Centre d'Etudes et de Conservation des Oeufs et du Sperme humain.

The French CECOS Federation collates the results of its 22 sperm banks and provides annual reports on their activity. These records allow studies on many different aspects; annual nationwide evaluation, matching of donors and recipients, follow-up of pregnancies, research into artificial procreation, and natural fertility. Risk of transmitted hereditary disease is minimised by genetic screening which establishes the genealogy of donor candidates and includes karyotyping and other biological investigations when a particular risk is suspected. The real risk of consanguinity is very small. Limiting the number of children born from a given sperm donor can be defined at the local level.

Influence of oxygen tension on reactive oxygen species production and human sperm function.

Human spermatozoa were exposed to the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) to stimulate endogenous production of reactive oxygen species (ROS). They were incubated under a gas phase of 5% CO2/90% N2/5% O2, or 5% CO2/95% air (20% O2) to investigate whether a lower than atmospheric oxygen tension in the gas phase of the incubator limited the endogenous production of ROS by human spermatozoa and thus was able to reduce the cytotoxic effects of ROS on sperm function. Exposure of human spermatozoa or exogenous NADPH induced an 8-fold higher production of superoxide anion under ambient vs. low oxygen tension. This marked difference in the stimulation of superoxide anion generation was associated with significantly different sperm motility parameters, according to the oxygen tension in the gas phase of the incubator. Whereas under 5% oxygen the percentage of motile spermatozoa was unaffected by the presence of NADPH, all of the motility parameters recorded under an atmosphere of 5% CO2 in air were dramatically affected, not only compared to their respective controls, but also compared to the motility parameters observed under low oxygen tension. The presence of superoxide dismutase plus catalase protected spermatozoa against the toxic effects of NADPH, confirming a cause/effect relationship between the increased superoxide production and reduced sperm function. Even at a concentration of NADPH which did not alter the percentage of motile spermatozoa, hyperactivated motility and acrosome reaction were significantly lower under an atmosphere of 5% CO2 in air compared to a gas phase of 5% CO2/90% N2/5% O2. These results suggest that there is an advantage in using 5% O2 rather than 20% O2 in the gas phase of the incubator to prevent the excessive production of ROS by spermatozoa and related alterations of sperm functions. This may be of clinical value in fertilization programmes.

Reactive oxygen species and human spermatozoa: physiology and pathology.

The role of reactive oxygen species (ROS) in the pathophysiology of human sperm function has been emphasized in recent years. ROS production in semen has been associated with loss of sperm motility, decreased capacity for sperm-oocyte fusion and loss of fertility. There is a current presumption that the most prolific source of ROS in sperm suspensions is an NADPH oxidase located in leukocytes or in spermatozoa which produces superoxide which is further converted to peroxide by the action of superoxide dismutase. Hydrogen peroxide has been recognized as the most toxic oxidizing species for human spermatozoa, which are very sensitive to lipid peroxidation owing to the high content of polyunsaturated fatty acids in their plasma membrane, though this is not the sole mechanism by which sperm function might be impaired by ROS. Although the excessive production of ROS is detrimental to human spermatozoa, there is a growing body of evidence which suggests that ROS are also involved in the physiological control of some sperm functions. This review focuses on the nature and source of the ROS generated by human spermataozoa as well as their operational mechanisms and their effects, which may be detrimental or beneficial.