PROK1 Level in the Follicular Microenvironment: A New Noninvasive Predictive Biomarker of Embryo Implantation.

CONTEXT: Prokineticin 1 (PROK1), also called endocrine gland-derived vascular endothelial growth factor, is a well-established regulator of endometrial receptivity and placental development. However, its clinical usefulness as a noninvasive predictive biomarker of embryo implantation is yet to be validated. OBJECTIVE: The main objective of this article was to determine the relationship between PROK1 levels in the follicular fluid (FF) and fertilization culture media (FCM) and the reproductive outcome in patients who received a first conventional in vitro fertilization-embryo transfer. The secondary objective was to characterize the expression of PROK1 and its receptors (PROKRs) in the human follicular microenvironment. DESIGN AND SETTING: We conducted a prospective study between January 2013 and June 2015 at the University Hospital of Grenoble. PATIENTS: A total of 135 infertile in vitro fertilization patients and 10 women undergoing ovarian tissue cryopreservation were included. INTERVENTIONS: The PROK1 concentration was measured by ELISA in FF and FCM collected on the day of oocyte retrieval and the day of the oocyte denudation step, respectively. Follicular expression of the PROK1/PROKR system was determined by immunohistochemistry, RT-quantitative PCR, and ELISA. MAIN OUTCOME MEASURE: Assessment of the clinical pregnancy rates was the main outcome. RESULTS: FF and FCM PROK1 levels were significantly higher in the embryo implantation group (P < .001) and were predictive of subsequent embryo implantation (area under the receiver operating characteristic curve, 0.91 [95% confidence interval, 0.81-1.00], P = .001; and 0.88 [0.72-1.00], P = .001, respectively). FF and FCM PROK1 levels remain similar irrespective of the embryo morphokinetic parameters (P = .71 and P = .83, respectively). The PROK1/PROKR system is expressed during human folliculogenesis. CONCLUSIONS: PROK1 levels in FF and FCM could constitute new predictive noninvasive markers of successful embryo implantation in conventional in vitro fertilization-embryo transfer.

In cryptozoospermia or severe oligozoospermia is sperm freezing useful?

BACKGROUND: Intracytoplasmic Sperm Injection (ICSI) is an Assisted Reproduction Technique (ART) which offers the chance to conceive to patients presenting very low sperm counts (cryptozoospermia/severe oligozoospermia). Sperm freezing before the oocyte pick-up, can prevent from a lack of spermatozoa on the day of the ICSI. It can avoid the cancellation of the ICSI or the use of TESE (Testicular sperm extraction). The objective of this study was to analyse the practice of sperm freezing for these patients in our center over 8 years and the rate of use of these frozen sperms. We also compared the outcome of ICSIs with frozen versus ejaculated sperm. MATERIAL AND METHODS: We performed a retrospective epidemiological study between 2004 and 2011. We recruited all the patients having a sperm count below 1 Million/mL and who were waiting for their first ICSI attempt. RESULTS: 169 patients were recruited: 84 cryopreserved their sperm before the ICSI (secured ICSI) while 85 did not (non-secured ICSI). Both groups were split in cryptozoospermia (<10(3) spermatozoa/ml): 19 and 17 patients respectively, very severe oligozoospermia (10(3)-10(5)/ml): 37 and 13 patients, and severe oligozoospermia (10(5)-10(6)/ml): 28 and 55 patients. The part of secured ICSI significantly increased from 29% during 2004-2007 to 74% during 2008-2011(p = 0.0029) and the frozen sperm was used in 5.9% of the cases. Median age was significantly higher in the non secured ICSI group (33.57 vs 35.52 for men, p = 0.0069 and 30.45 vs 32.26 for women, p = 0.025) but no significant difference was found in the outcome of the ICSI between frozen-thawed sperm and fresh ejaculated sperm. CONCLUSION: Sperm freezing before ICSI for severe oligozoospermic and cryptozoospermic patients significantly increased in our practice but the rate of use remain very low. This encourages to define more accurate criteria leading to sperm freezing.

CONTEXTE: L'ICSI (Intracytoplasmic Sperm Injection) permet aux patients porteurs d'une oligozoospermie sévère ou d'une cryptozoospermie d'espérer concevoir un enfant. Ces patients courent parfois le risque de survenue d'une azoospermie constatée le jour de l'ICSI. L'organisation préalable d'une ou plusieurs congélations spermatiques peut permettre de prévenir ce risque. L'objectif de cette étude était d'évaluer cette pratique de congélation spermatique de sécurité sur une période prolongée dans notre centre et d'évaluer le taux d'utilisation de ces spermes congelés et les chances de grossesse au cours des ICSI, qu'elles soient faites avec les spermatozoïdes frais ou congelés et selon la gravité de l'oligozoospermie. MATERIEL ET METHODES: Il s'agit d'une étude rétrospective monocentrique descriptive de 2004 à 2011. Les critères d'inclusion étaient une numération spermatique inférieure à 1 Million/mL pour une première tentative d'ICSI. RESULTATS: 169 patients ont été recrutés: 84 ont bénéficié de congélation de spermatozoïdes avant l'ICSI (ICSI sécurisée) et 85 patients n'en ont pas bénéficié (ICSI non sécurisée). Au sein de ces deux groupes, les patients ont été répartis en 3 sous-groupes en fonction de leur concentration spermatique: cryptozoospermie (<103 spermatozoïdes/ml), 19 et 17 patients, oligozoospermie très sévère (103­105/ml), 37 et 13 patients, et oligozoospermie sévère (105­106/ml): 28 et 55 patients. Durant la période analysée nous avons observé une augmentation significative du nombre de congélations spermatiques préalables à une ICSI, passant de 29% des tentatives pour 2004­2007 à 74% des tentatives pour 2008­2011 (p = 0.0029. Seulement 5,9% des patients ont utilisé les spermatozoïdes congelés. L'âge moyen des patients s'est avéré significativement plus élevé dans le groupe des ICSI sans congélation spermatique que avec congélation spermatique (33.57 vs 35.52 chez les hommes, p = 0.0069 et 30.45 vs 32.26 chez les femmes, p = 0.025) sans qu'il y ait de différence significative dans l'issue des tentatives d'ICSI. CONCLUSION: La congélation spermatique avant ICSI dans le cas des oligozoospermies sévères et cryptozoospermies est devenue presque systématique dans notre pratique. Pour autant l'utilisation de ces spermatozoïdes congelés demeure très peu fréquente. Ceci nous engage à poursuivre l'étude de ces situations afin de définir des critères cliniques et/ou biologiques justifiant ces congélations souvent itératives.