The aim of the present study was to evaluate the effect of activin A on the activation of in vitro folliculogenesis of human ovarian tissues from transgender men with or without our new compartmented chitosan hydrogel microbioreactor (“three-dimensional (3D)-structure”) enabling a three-dimensional tissue culture. Five fresh ovarian human tissues were cultured in vitro for 20 or 22 days in four groups with 100 ng/mL activin A or without activin A during the last six to eight days of culture, and within a 3D-structure or without the 3D-structure in standard conditions. Follicular density and quality were evaluated, and follicular diameters were measured. Estradiol secretion was quantified. Proliferation and apoptosis through immunostaining were also performed. The proportion of primordial follicles was significantly reduced, and the proportion of primary and secondary follicles was significantly increased in all four groups (p < 0.001). Tertiary follicles were observed in the four culture groups. Activin A supplementation did not significantly affect the follicular density, follicular quality, follicular growth, or estradiol secretion (p > 0.05). The 3D-structure increased the density of primary follicles and decreased the estradiol secretion (p < 0.001). Follicular proliferation was significantly lower in the 3D-structure group compared to the non-3D-structure group (p = 0.008). Regarding follicular apoptosis, it was significantly higher in the activin group compared to the non-activin group (p = 0.006). Activin A did not seem to play a key role in the in vitro folliculogenesis activation in our culture conditions. However, the results may indicate that the 3D-structure could be more physiological and could prevent a detrimental in vitro folliculogenesis flare-up.
Background: The aim of the present study was to evaluate the effect of Activin A on the activation of in vitro folliculogenesis of human ovarian tissues with or without our new three-dimensional structure (3D-structure). Methods: Five fresh ovarian human tissues were cultured in vitro in 4 groups with 100ng/mL Activin A or without Activine A and within or without a 3D-structure for 20 or 22 days of in vitro culture. Follicular density and quality were evaluated, and follicular diameters were measured. Estradiol secretion was quantified. Proliferation and apoptosis through immunostaining were performed.Results: The proportion of primordial follicles was significantly reduced, and the proportion of primary and secondary follicles was significantly increased in all four groups (p<0.001). Antral cavities were observed in the four culture groups. Activin A supplementation did not significantly affect the follicular density, follicular quality, follicular growth, or estradiol secretion (p>0.05). The 3D-structure increased the density of primary follicles and decreased the estradiol secretion (p<0.001). Tissular proliferation was significantly lower in the 3D-structure group compared to the non-3D-structure group (p=0.008). Regarding tissular apoptosis, it was significantly higher in the Activin group compared to the non-Activin group (0.006). Conclusion: The presence of Activin A did not seem to play a key role in in vitro folliculogenesis activation. However, the results may indicate that the 3D-structure could be more physiological and could prevent a pejorative in vitro folliculogenesis flare-up.
Study question Does the use of testicular spermatozoa from non-azoospermic infertile men after failed IVF with ejaculated spermatozoa increases the pregnancy rate? Summary answer Depending on semen characteristics and obtained embryos number at the previous IVF failure with ejaculated spermatozoa, it seems licit to propose ICSI with testicular spermatozoa. What is known already Ejaculated spermatozoa are generally considered to possess better fertilisation potential than testicular spermatozoa as their maturation has been completed. However, it has been shown that in selected cases, the use of testicular spermatozoa from non-azoospermic infertile men resulted in a higher implantation and pregnancy rate than the use of ejaculated spermatozoa. One of the reasons for this result is a higher DNA fragmentation index (DFI) in ejaculated spermatozoa compared to testicular spermatozoa. This increase in DFI would negatively affect embryo development and implantation. Study design, size, duration A retrospective matched cohort study using propensity score matching (PSM) analysis was performed. After an IFV failure (cycle_1), IVF with ejaculated or testicular spermatozoa was performed (cycle_2). Female age, female BMI, IVF rank, stimulation protocol, gonadotropin total quantity, treatment duration, punctured oocytes number, and mature oocytes punctured were included in PSM. The matching was performed for IVF performed in cycle_1 for cases and controls identified in cycle_2. 26 couples were included between 01/01/2012 and 30/09/2021. Participants/materials, setting, methods Among the 126 couples, for 63 couples, the cycle_2 was performed with ejaculated spermatozoa (controls) and for 63 couples with testicular spermatozoa (cases). Mixed logistic regression was used to compare outcomes of cases versus controls groups. The cycle_1 have allowed finding the prognostic factors to propose a cycle_2 ICSI with testicular spermatozoa in case of cycle_1 IVF failure. The study outcomes were the pregnancy rate (PR) and the cumulative pregnancy rate (CPR). Main results and the role of chance In cycle_1, no difference was observed for the parameters included in PSM. The DFI was higher in case group (13.7% ± 10.5% versus 9.3% ± 4.4%, p < 0.05), no difference was observed for fertilization rate, blastulation rate and frozen embryo rate. However, the PR was higher in case group (22.2% versus 0.0%, p < 0.001), the same result was found for the CPR (25.4% versus 6.3%, p < 0.001). The main prognostics factors to propose a cycle_2 ICSI with testicular spermatozoa after cycle_1 IVF failure with ejaculated spermatozoa: teratozoospermia or cryptozoospermia in cycle_1 (OR = 6.0 [1.2 ; 31.1], p < 0.05), a number of obtained embryos greater than 7 in cycle1 (OR = 5.3 [1.5 ; 18.0], p < 0.01), independently of male age in cycle_1 (OR = 2.3 [0.8 ; 6.5], p > 0.05 Limitations, reasons for caution The main limitations of the current study is being retrospective rather than prospective randomized. However, the used of the propensity score to perform a case-control study allow this study to be more reliable and to obtain results corresponding to real life. Wider implications of the findings This study is in favour of using testicular spermatozoa after IVF failure with ejaculated spermatozoa when the semen morphology is altered or in case of cryptozoospermia, and when at least 8 embryos was obtained in the previous IVF procedure Trial registration number not applicable
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