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6 results on '"Cyrielle Fournier"'

1. A New Bioreactor to Promote Human Follicular Growth with or without Activin A in Transgender Men

Author

Cynthia Jovet, Eloïse Fraison, Jacqueline Lornage, Nicolas Morel Journel, Antoine Gavoille, Laurent David, Alexandra Montembault, Cyrielle Fournier, Bruno Salle, and Elsa Labrune

Subjects
in vitro folliculogenesis, bioreactor, activin A, follicle growth, ovarian tissue culture, Reproduction, QH471-489
Abstract

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.

Published
2023
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2. Development and Survival of Human Ovarian Cells in Chitosan Hydrogel Micro-Bioreactor

Author

Elsa Labrune, Cyrielle Fournier, Benjamin Riche, Laurent David, Alexandra Montembault, Sophie Collardeau-Frachon, Mehdi Benchaib, Jacqueline Lornage, Jean Iwaz, and Bruno Salle

Subjects
chitosan, hydrogel, folliculogenesis, bioreactor, tissue culture technique, Medicine (General), R5-920
Abstract

Background and Objectives: To test the long-term ability of human ovarian cortex cells to develop in unconventional culture conditions. Materials and Methods. Ovarian cortex cells from fetuses aged 23 to 39 weeks gestation were cultured for 90 days in hollow chitosan hydrogel micro-bioreactors and concurrently in traditional wells. Various cell-type counts were considered. Results: With intact follicles as a denominator, the percentage of growing intact follicles at Day 0 varied widely between ovaries (0 to 31.7%). This percentage tended to increase or stay relatively constant in bioreactor as in control cultures; it tended more toward an increase over time in bioreactor vs. control cultures. Modeled percentages showed differences (though not significant) in favor of bioreactor cultures (16.12% difference at D50 but only 0.12% difference at D90). With all follicles present as a denominator, the percentage of growing primary and secondary follicles at D0 varied widely between ovaries (0 to 29.3%). This percentage tended to increase over time in bioreactor cultures but to decrease in control cultures. Modeled percentages showed significant differences in favor of bioreactor cultures (8.9% difference at D50 and 11.1% difference at D90). At D50 and D90, there were only few and sparse apoptotic cells in bioreactor cultures vs. no apoptotic cells in control cultures. Conclusions: Over three months, bioreactor folliculogenesis outperformed slightly traditional culture. This is an interesting perspective for follicle preservation and long-term toxicological studies.

Published
2022
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3. Human ovarian cryopreservation: vitrification versus slow freezing from histology to gene expression

Author

Jacqueline Lornage, Nicolas Morel-journel, Pauline Jaeger, Cyrielle Fournier, Mehdi Benchaib, Bruno Salle, Eloise Fraison, Claire Santamaria, and Elsa Labrune

Subjects
Slow freezing, Andrology, Reproductive Medicine, Gene expression, Ovarian cryopreservation, Obstetrics and Gynecology, Vitrification, Histology, General Medicine, Biology
Abstract

Background: Cryopreservation of ovarian tissue is one of the strategies offered to girls and women needing gonadotoxic treatment, as a means of preserving their fertility. There are two methods of ovarian tissue cryopreservation: slow freezing, the reference method, and vitrification, an alternative method. The aim of the present study was to evaluate which of the two is the best method for human ovarian tissue cryopreservation. Each ovary was divided into 3 groups: Fresh, Slow freezing and Vitrification. In each group a histological study to evaluate follicular density and quality; and an evaluation of 6 gene expression (CYP11A, STAR, GDF9, ZP3, CDK2, CDKN1A) were performed. Results: We observed no significant difference in follicular density within these 3 groups. Slow freezing altered the pool of primordial follicles compared to the Fresh tissue (31.8% vs 55.9%, p = 0.046, respectively). The expression of genes involved in steroidogenesis varied after cryopreservation compared to the fresh group; CYP11A was under-expressed in both freezing groups compared to the fresh group and significantly under-expressed in the slow freezing group (p = 0.01), STAR was over-expressed in the slow freezing group and significantly under-expressed in the vitrification group (p = 0.01). Regarding the expression of genes involved in cell cycle regulation, CDKN1A was significantly under-expressed in both freezing groups (slow freezing: p = 0.0008; vitrification: p = 0.03) compared to the fresh group. Conclusion: Vitrification had no effect on the histological quality of the follicles at any stage of development compared to Fresh tissue. There was no significant difference in gene expression between the two techniques.

Published
2022

4. Cellular and Molecular Impact of Vitrification Versus Slow Freezing on Ovarian Tissue

Author

Jacqueline Lornage, Bruno Salle, Cyrielle Fournier, M. Benchaib, Claire Santamaria, Muriel Rabilloud, Elsa Labrune, and Pauline Jaeger

Subjects
0206 medical engineering, Biomedical Engineering, Medicine (miscellaneous), Apoptosis, Bioengineering, Ovary, 02 engineering and technology, Tissue Culture Techniques, Andrology, 03 medical and health sciences, Ovarian Follicle, Freezing, Gene expression, medicine, Animals, Vitrification, 030304 developmental biology, Cryopreservation, Slow freezing, 0303 health sciences, Sheep, Chemistry, Gene Expression Profiling, Ovarian tissue, Histology, 020601 biomedical engineering, medicine.anatomical_structure, Gene Expression Regulation, Female, Tissue Preservation
Abstract

Objective: To evaluate a vitrification protocol from histology to gene expression to slow freezing. Methods: Ovaries from 12 prepubertal ewes. The same ovary was cut into fragments, studied fresh, ...

Published
2020

5. A New Bioreactor to Promote Follicular Growth With or Without Activin A

Author

Cynthia Jovet, Jacqueline Lornage, Antoine Gavoille, Elsa Labrune, David Laurent, Nicolas Morel-journel, Eloise Fraison, Alexandra Montembault, Bruno Salle, and Cyrielle Fournier

Subjects
Activin a, endocrine system, Chemistry, Follicular phase, Bioreactor, Cell biology
Abstract

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 (p0.05). The 3D-structure increased the density of primary follicles and decreased the estradiol secretion (pConclusion: 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.

Published
2021

6. The impact of histones linked to sperm chromatin on embryo development and ART outcome

Author

M. Benchaib, Jacqueline Lornage, G. Soignon, S. Giscard D’estaing, Elsa Labrune, Jean-François Guérin, and Cyrielle Fournier

Subjects
Adult, Male, 0301 basic medicine, Pregnancy Rate, Reproductive Techniques, Assisted, Urology, Endocrinology, Diabetes and Metabolism, medicine.medical_treatment, Embryonic Development, macromolecular substances, Biology, Cohort Studies, Histones, Andrology, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Pregnancy, medicine, Humans, Prospective Studies, Protamines, Blastocyst, Prospective cohort study, 030219 obstetrics & reproductive medicine, Assisted reproductive technology, Embryogenesis, Embryo, medicine.disease, Spermatozoa, Sperm, Chromatin, Pregnancy rate, 030104 developmental biology, medicine.anatomical_structure, Reproductive Medicine, Female
Abstract

The purpose of this study was to investigate the relationship between the proportion of sperm chromatin linked to remaining histone and assisted reproductive technology (ART) outcome. A prospective cohort study was performed on couples undergoing ART process at the Department of Reproduction Medicine (HFME, Bron, France). The histone-to-protamine ratio (HPR) was measured using the method described by WykesKrawetz (2003) J Biol Chem 278, 29471. The correlations with sperm DFI, blastocyst formation, pregnancy rate, and delivery rate were investigated. A total of 291 ART cycles were included (42 c-IVF and 249 ICSI procedures): 3870 oocytes were punctured and 2211 embryos were obtained, among which 507 were transferred and 336 frozen. The mean HPR was 18.9%. A significant negative correlation was found between HPR and DFI (r = -0.12, p 0.05). Regarding the type of ART procedure (c-IVF or ICSI), the same kind of relationship between HPR and ART parameters was observed. Regardless of the type of ART procedure used, when the HPR was within the range [6%; 26%], the blastocyst formation rate was higher: 87.8% vs. 71.2% (HPR6%; p 0.01) and 74.6% (HPR26%; p 0.01). The highest delivery rate (DR; 24.5%) was obtained for HPR within the range [6%; 26%]; DR was 21.9% for HPR6% and 18.3% for HPR26%; however, the differences were not statistically significant. The procedure described in this study seems to be a reliable evaluation of the HPR. The HPR parameter seems to be correlated to embryonic development up to the blastocyst stage, but its involvement in clinical pregnancy/delivery could not be confirmed. HPR should be further investigated for confirming the relationship with blastocyst formation. After this, the next step will be to investigate the etiologies of HPR alterations for improving the sperm nucleus quality for increasing the chance of pregnancy.

Published
2018

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