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Background & Aim Mesenchymal Stromal Cells (MSCs) are involved in several clinical trials in the field of Advanced Therapeutic Medicinal Products (ATMPs) for their properties and therapeutic potential. However, the production of MSCs is currently done in multi-plates culture chambers (CellSTACK, Corning) that only allow the production of a limited number of cells and it's time consuming. This therefore represents a constraint, especially for allogeneic uses. Now, the major challenge is to increase this production capacity. The use of a bioreactor would overcome this problem. The Xpansion® bioreactor (Pall) is composed of a controller and single-use bioreactors. Similar in concept to CellSTACK, the Xpansion bioreactors are composed of stacked cell culture plates that are sold in four variations: 10, 50, 100, and 200. Although the plastic of the bioreactor plates is identical to those of the CellSTACK, the feasibility and equivalence of quality and expansion of the MSCs produced using the Xpansion bioreactor had to be demonstrated. Methods, Results & Conclusion Preliminary tests were carried out on the smallest model of the Xpansion bioreactor product line: The XP10 (10 culture plates). All experiments were performed in parallel with a CellSTACK manipulation. The tests were performed using MSCs at passage 0 (from bone marrow or adipose tissue) or raw materials. Quantitative and qualitative tests were carried out at the end of the culture. From raw material or MSCs, cell densities obtained after 6 days of culture are equivalent in both the Xpansion bioreactor and CellSTACK, demonstrating similar cell growth. The average cell expansion rates in the Xpansion (XP10) bioreactor and CellSTACK are 24.39 times and 21.69 times (n=3), respectively. The flow cytometry analysis showed that the phenotype and viability of the MSCs from the bioreactor were not altered by this culture method. Preliminary tests on the Xpansion bioreactor have demonstrated the feasibility of MSCs cell expansion without affecting the quality of the finished product compared to the current method in CellSTACK. In addition, using a bioreactor allows the production of cells in large quantities by managing only one culture chamber while reducing the risk of contamination and operation time. Additionally, using a bioreactor improves monitoring of culture parameters and scalability. Manufacturing MSCs in this bioreactor would make the graft of a larger number of patients from the same initial collection possible.

Journal de Gynecologie Obstetrique et Biologie de la Reproduction - Vol. 33 - N° 6-C1 - p. 518-524

Le travail rapporte l’experience parisienne du diagnostic genetique pre-implantatoire (DPI). 59 couples ont ete pris en charge de janvier 2000 a juillet 2001 au cours de 71 cycles. Les ovocytes recueillis ont ete insemines par injection intracytoplasmique de spermatozoide et les embryons biopsies au troisieme jour de developpement. L’analyse genetique a ete effectuee le jour de la biopsie et la majorite des transferts embryonnaires effectues le quatrieme jour post fecondation. Au cours des 71 cycles, 872 ovocytes ont ete recueillis dont 731 etaient compatibles avec une injection intracytoplasmique de spermatozoide. Parmi les 505 embryonds obtenus, 421 embryons ont ete biopsies et un resultat genetique a ete obtenu pour 312 (74%) d’entre eux. 127 embryons ont ete transferes au cours de 58 transferts. 18 grossesses biologiques et 12 cliniques ont ete obtenues (7 uniques, 4 gemellaires et une triple). 16 enfants sont nes. Le DPI prend desormais sa place parmi les choix proposes aux couples a risque de transmission de maladie genetique de particuliere gravite et incurable au moment du diagnostic.

Objective To assess intrauterine levels of leukemia inhibitory factor (LIF) by uterine flushing at the time of egg retrieval and to confirm that the procedure has no detrimental effect on pregnancy rates. Design Prospective study. Setting Assisted reproductive unit of a university hospital. Patient(s) Uterine flushing was performed in 148 IVF patients. The first 100 patients were compared with a matched control group. Intervention(s) Uterine flushing at the time of egg retrieval. Main outcome measure(s) IVF-ET results, pregnancy rates, and intrauterine LIF levels. Result(s) Pregnancy rates were not different in the group of patients with (27%) or without uterine flushing (28%). Leukemia inhibitory factor was detected in 60 patients (46%). Pregnancy rates did not differ between patients' detectable LIF and those in whom LIF was undetectable. Mean levels of LIF were 30.1 ± 49.3 pg/mL and 28.6 ± 51.2 pg/mL in pregnant and nonpregnant patients respectively. Conclusion(s) The flushing procedure at the time of egg retrieval did not adversely affect pregnancy rates. Leukemia inhibitory factor was detected in 46% of patients at the time of egg retrieval, but no correlation were observed with better pregnancy rates in patients with detectable LIF. Mean LIF levels did not differ in pregnant and nonpregnant women. Access to endoluminal secretions of the endometrium during IVF-ET may represent a new research in human implantation.

Resume Objectifs. – Presenter les alternatives chirurgicales au traitement du syndrome des ovaires polykystiques (SOPK). Patientes et methodes. – Soixante et onze patientes traitees par drilling ovarien par cœlioscopie et 47 par hydrolaparoscopie. Le critere principal d’inclusion etait la resistance a six mois de traitement par citrate de clomifene chez des patientes oligo-amenorrheiques. Resultats. – Apres une revue des principaux resultats des series chirurgicales publiees de drilling ovarien, nous retrouvons 73 % de grossesse apres traitement cœlioscopique et 61 % apres hydrolaparoscopie. Discussion et conclusion. – Les resultats sur la fertilite des drillings ovariens en font une alternative aux stimulations avec insemination intra-uterine apres echec du citrate de clomifene et/ou de la metformine. Le traitement chirurgical limite le risque de grossesses multiples et de fausses couches. L’hydrolaparoscopie apparait aussi efficace que la cœlioscopie et moins morbide chez des patientes souvent obeses.

Background: Pregnancy occurring after pelvic arterial embolization for postpartum hemorrhage is rare. The harmlessness of this procedure for subsequent pregnancies is poorly documented. Materials and methods: Eight pregnant women with a history of arterial embolization were followed up with fetal growth and Doppler assessment. Results: Fetal growth and umbilical Doppler findings remained normal in all cases. Conclusion: Embolization, a life-saving and fertility-preserving procedure, does not seem to affect subsequent pregnancies.

RESUME Le travail rapporte l’experience parisienne du diagnostic genetique preimplantatoire (DPI). Cinquante-neuf couples ont ete pris en charge de janvier 2000 a juillet 2001 au cours de 71 cycles. Les ovocytes recueillis ont ete insemines par injection intracytoplasmique de spermatozoide et les embryons biopsies au troisieme jour de leur developpement. L’analyse genetique a ete effectuee le jour de la biopsie et la majorite des transferts embryonnaires ont eu lieu le quatrieme jour post fecondation. Au cours des 71 cycles, 872 ovocytes ont ete recueillis dont 731 etaient compatibles avec une injection intracytoplasmique de spermatozoide. Parmi les 505 embryons obtenus, 421 ont ete biopsies et un resultat genetique a ete obtenu pour 312 (74 %) d’entre eux. Cent vingt-sept embryons ont ete transferes au cours de 58 transferts. Dix-huit grossesses biologiques et 12 cliniques ont ete obtenues (7 uniques, 4 gemellaires et une triple). Seize enfants sont nes. Le DPI prend desormais sa place parmi les choix proposes aux couples a risque de transmission d’une maladie genetique de particuliere gravite et incurable au moment du diagnostic.

Patients with a history of repeated and unexplained implantation failure after IVF-embryo transfer and a high uterine natural killer cell count during estrogen-progestin treatment may benefit from controlled natural IVF-ET.

Journal de Gynecologie Obstetrique et Biologie de la Reproduction - Vol. 33 - N° 1-C2 - p. 33-35

To evaluate the efficacy of semi natural cycle as option of treatment for the patients who have a poor prognosis to controlled ovarian hyperstimulation (COH).Prospective study. Sixty-six patients with an ovulatory cycle who presented either altered ovarian status (AOS) or implantation failure (IMF) and had carried out a total of 133 semi natural IVF cycles were included. Once follicular dominance was established, the growth of this follicule was controlled by administration of the GnRH antagonist with exogenous gonadotrophins.Groups AOS (n = 47) and IMF (n = 19) were similar with regard to oocyte pickup rate (81.2% and 81.1%), the oocyte recovery rate (61.4% and 64.8%) and clinical pregnancies per oocyte pickups rate (15.4% and 16.6%), respectively.The semi natural cycle appears as option of treatment for patients who have a poor prognosis for successful in vitro fertilization.

Study Objective To evaluate fertiloscopy ovarian drilling with bipolar energy in women with clomiphene citrate-resistant polycystic ovary syndrome (PCOS). Design Prospective study (Canadian Task Force classification II). Setting University teaching hospital and private clinic. Patients Eighty women with clomiphene citrate-resistant PCOS. Intervention Operative transvaginal fertiloscopy with a coaxial bipolar electrode. Measurements and Main Results During a mean follow-up of 18.1 months (± 6.4), 73 women (91%) recovered regular and ovulatory cycles. The cumulative pregnancy rate was 60% (44/73) for spontaneous and stimulated cycles, with 39.7% (29/73) imputed to drilling alone. The mean time to conceive was 3.9 months (range 1–11.8). There were eight miscarriages (18%), and no ectopic pregnancies or multiple pregnancy. No complications occurred. Conclusion Ovarian drilling by transvaginal fertiloscopy with bipolar electrosurgery appears to be an effective minimally invasive procedure in patients with PCOS resistant to clomiphene citrate.

Although controlled ovarian hyperstimulation (COH) has proved more effective than in vitro fertilization (IVF) done on natural cycles, poor responses related to ovarian status defects are not rare and may account for cancellation rates as high as 24% in IVF-embryo transfer (ET) cycles. Both women with low ovarian reserve and those responding poorly to previous ovarian stimulation may be affected. This prospective study was an attempt to determine whether seminatural cycles are a reasonable approach to patients with ovarian aging. A total of 158 cycles were monitored in 75 women. Participants were infertile women less than 38 years of age who had ovulatory cycles every 25 to 35 days and ovarian aging, defined as low ovarian reserve and characterized by high levels of follicle-stimulating hormone by cycle day 3, high estradiol and/or low inhibin B levels, and/or previous cycle cancellations because of a poor ovarian response to COH. Up to 3 cycles were offered. The dominant follicle, selected from cycle day 8 onward, was monitored by ultrasonography and hormone measurements. When it appeared, as evidenced by a serum estradiol exceeding 100 pg/mL and a diameter of more than 12 mm, patients received a gonadotropin-releasing hormone (GnRH) antagonist followed by human menopausal gonadotropin. Oocyte pickup was done 36 hours after administration of human chorionic gonadotropin. Embryo transfers were carried out 2 days after oocyte retrieval. The luteal phase was supported by micronized progesterone. The cancellation rate for 158 cycles was 17.7%. Oocyte pickups were done in 75.3% of cycles, and 91 mature oocytes were retrieved (57.6%). The rate of embryo transfer was 42.4%. Nineteen clinical pregnancies occurred, 5 of which resulted in spontaneous abortion. Seven of the pregnancies occurred in the first treatment cycle. The cumulative rate of pregnancy per patient after 3 treatment cycles was 35.2%. The investigators concluded that IVF with a seminatural cycle protocol is a useful alternative to COH for those who fail to respond well. Candidates include women who require IVF, those with endocrinologic evidence of ovarian aging, and those having a history of one or 2 canceled COH cycles. Better results may be expected as the dose and duration of treatment with a GnRH antagonist are appropriately modified and as the oocyte pickup technique improves.

BACKGROUND: Most implantation failures after successful in vitro fertilization-embryo transfer (IVF-ET) result from inadequate uterine receptivity. There is currently no way to predict this receptivity. METHODS: We investigated whether the detection of interleukin-(IL)18 by ELISA in uterine luminal secretions might predict implantation failure. Secretions of 133 patients enrolled in our IVF-ET program were sampled by uterine flushing immediately before oocyte retrieval. We assessed the following outcomes: pregnancy rate, multiple pregnancy rate, and implantation rate per embryo transferred. RESULTS: Interleukin-18 was detected in the flushing fluid of 38 patients (28.6%). Although the two groups were comparable for all other characteristics (age, etiology, ovarian reserve, number of embryos transferred, quality of embryos), all outcome variables differed significantly. The pregnancy rate was 37.9% in the IL-18 - ve group and 15% in the IL-18 + ve group, the multiple pregnancy rate 27.7% and 0%, and the implantation rate per embryo transferred 19.4% and 6.7% (all comparisons, P = 0.02). Only embryos meeting good quality criteria were transferred to 65 patients: 50 IL-18 - ve and 15 IL-18 + ve. The pregnancy rate was 51 % for the IL-18 - ve group and 20% for the IL-18 + ve group, the multiple pregnancy rate 36% and 0.0%, respectively, and the implantation rate 29% and 8.3% (P = 0.02). CONCLUSION: This non-invasive and simple method predicted inadequate uterine receptivity, independent of embryo quality.

Since the introduction of IVF treatments, natural cycle IVF (nIVF) has been largely replaced by IVF with ovarian stimulation to obtain multiple oocytes. Failure to anticipate the retrieval of more than two oocytes often results in cancellation of the cycle. However, nIVF has several advantages. It is associated with a close to zero multiple pregnancy rate, and a zero risk of ovarian hyperstimulation syndrome. Per cycle, nIVF is less time consuming, physically and emotionally less demanding for patients, and cheaper than stimulated IVF, but also less effective. GnRH antagonists have improved the outcome of nIVF cycles. They avoid unexpected ovulation and frequent cancellations. In selected cases, especially in women with implantation failure or with poor ovarian response, favorable results can be obtained from nIVF cycles, despite the replacement of a single embryo. Poor results are obtained in women over 38 years.

Show the different surgical approaches for the treatment of polycystic ovarian syndrome (SOPK).Seventy-one patients treated by ovarian drilling by laparoscopy and 47 by hydrolaparoscopy. The main inclusion criterion was the absence of response after 6 months of stimulation by clomifene citrate for oligo-amenorrheic patients.After a review of surgical series published of ovarian drilling, we found 73% pregnancy rate after laparoscopic treatment and 61 % after hydrolaparoscopy.Ovarian drilling appears as an alternative to ovarian hyperstimulation with intra-uterine insemination after failure of clomifène citrate and/or metformine. Surgical treatment decreases multiple pregnancies and miscarriages. Hydrolaparoscopy is more efficient than laparoscopy with less morbidity in particular in obese patients.

The contractile activity of the nonpregnant uterus plays an important role in the human reproduction process. Noticeable variation in uterine contractility occurs during the menstrual cycle to meeting some physiologic requirements of the female reproductive system. During the follicular phase, stimulation of uterine contractions by estrogens fosters sperm transport toward the fertilization site. After ovulation, contractility decreases in response to progesterone, a phenomenon that is probably involved in the embryo implantation mechanism. The comprehension of uterine contractility regulation and physiologic roles has been considerably extended during recent years by the development of direct and noninvasive assessment tools, in particular, ultrasound. Today, not only the artificial stimulation of uterine contractions, aiming at promoting sperm transport during the pre-ovulatory phase, but also their attenuation to provide optimum conditions for embryo implantation during the luteal phase, represent innovating, promising issues in the optimization of assisted reproduction treatments.

Introduction. The TVT procedure has been changed the surgical treatment of stress urinary incontinence (SUI). The aim of this study was to collect the patient's point of view.andThe TVT procedure was performed to treat SUI on 144 consecutive women in our institution between May 1998 and December 2000. We have sent a postal questionnaire to all patients in order to determine subjective results on SUI and post-operative voiding difficulties.We have obtained 124 responses (86.1% of patients). The median follow-up was 20 months +/- 12 (range 8 to 38 months). The patients were "very satisfied" in 56.5% (n=70), "satisfied" in 34.6% (n=43) and "unsatisfied" in 8.9% of cases (n=11). The functional discomfort about SUI was, on a 0 to 10 visual analog scale, 7.5 +/- 2 preoperatively and 2.2 +/- 2.8 post-operatively. Only 13.7% of patients have SUI postoperatively, and then 86.3% are considered cured. Nevertheless, voiding difficulties are common after this procedure with 31.5% of patients complaining about dysuria. On 96 patients who had had mixed incontinence preoperatively, 45.8% (n=44) complaining about persistent urge symptoms. On 28 patients who had had pure SUI, 28.6% (n=8) complaining about de novo urge symptoms.The TVT procedure is an efficient treatment of stress urinary incontinence, but with numerous post-operative voiding difficulties. This conclusion justify a precise pre-operative information to the patients.

Background There is strong evidence that locally secreted cytokines control the implantation process and can cause implantation failure. Uterine flushing fluids were analysed to determine their concentrations of leukaemia inhibitory factor (LIF) and tumour necrosis factor (TNF). Methods and results We began by flushing the uterine cavities of 33 infertile patients on day 26 of two consecutive cycles. The concentrations of LIF (by enzyme-linked immunosorbent assay) and TNF (by bioassay) were significantly correlated during these cycles (r = 0.762, P = 0.0001 and r = 0.822, P = 0.001 respectively) and hence reliable. Then, after a reference flushing of 30 infertile patients, we followed the outcome of their first consecutive cycle of ovarian stimulation, which preceded either IVF or intrauterine insemination. A total of 10 patients became pregnant. The median concentration of LIF was 0 pg/ml (range: 0-177) and of TNF was 0 U/ml (range: 0-6.17) among those who became pregnant, and 203 pg/ml (range: 0-1620) and 2.14 U/ml (range: 0-16) respectively among those who did not. The LIF concentration was significantly lower in the pregnant group (P = 0.0013). Conclusion A low concentration of LIF in the uterine flushing fluid at day 26 was predictive of subsequent implantation. Use of this procedure should increase the number of IVF attempts yielding successful pregnancies and also lead to corrective therapies.

Premature LH and progesterone surges are associated with different factors and hormonal modulators. The aim of the present study was (i) to investigate the clinical and laboratory factors and (ii) to highlight the importance of different stimulation protocols in associated premature LH and progesterone surges in intrauterine insemination (IUI) cycles. The study involved a retrospective investigation of 75 patients undergoing IUI for infertility treatment (135 IUI cycles) between 1996 and 2000, with initial serum LH concentrations10 mIU/ml during ovarian stimulation. Ultrasound characteristics, follicular sizes, serum oestradiol, progesterone and LH concentrations and ovarian stimulation protocols were measured. There was a wide range of oestradiol serum concentrations (93-2245 pg/ml) and follicular size (12-25 mm). In 49.6% of cycles, the dominant follicle was16.5 mm. Patients with2 follicles measuring15 mm had higher oestradiol serum concentrations (P = 0.008). Multiple regression analyses revealed no association between these variables and premature LH/progesterone surge. In conclusion, LH/progesterone surges cannot be predicted utilizing clinical parameters normally employed, e.g. ultrasound serum oestradiol assay or ovarian stimulation protocol. Patients with follicles14 mm or more and with high numbers of small follicles and high oestradiol are at risk of a spontaneous LH surge. These variables can be used to time the administration of GnRH antagonist administration until better predictive factors are demonstrated.

In France, all embryos obtained after ovum donation have to be frozen. We present a brief background on this policy and expose our results, then discuss the rationale of such a policy in order to upgrade knowledge on the mechanism of vertical HIV transmission.