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. 2024 May;309(5):2127-2136.
doi: 10.1007/s00404-024-07419-z. Epub 2024 Mar 12.

Survival and hormone production of isolated mouse follicles in three-dimensional artificial scaffolds after stimulation with bpV(HOpic)

Philip Keckstein  1 , Ralf Dittrich  2 , Nathalie Bleisinger  2 , Inge Hoffmann  2 , Matthias W Beckmann  2 , Albrecht Gebhardt  3 , Benjamin Schmid  4 , Simon Keckstein  5
Affiliations

Survival and hormone production of isolated mouse follicles in three-dimensional artificial scaffolds after stimulation with bpV(HOpic)

Philip Keckstein et al. Arch Gynecol Obstet. 2024 May.

Abstract

Purpose: To preserve fertility before gonadotoxic therapy, ovarian tissue can be removed, cryopreserved, and transplanted back again after treatment. An alternative is the artificial ovary, in which the ovarian follicles are extracted from the tissue, which reduces the risk of reimplantation of potentially remaining malignant cells. The PTEN inhibitor bpV(HOpic) has been shown to activate human, bovine and alpacas ovarian follicles, and it is therefore considered a promising substance for developing the artificial ovary. The purpose of this study was to examine the impact of different scaffolds and the vanadate derivative bpV(HOpic) on mice follicle survival and hormone secretion over 10 days.

Methods: A comparative analysis was performed, studying the survival rates (SR) of isolated mice follicle in four different groups that differed either in the scaffold (polycaprolactone scaffold versus polyethylene terephthalate membrane) or in the medium-bpV(HOpic) versus control medium. The observation period of the follicles was 10 days. On days 2, 6, and 10, the viability and morphology of the follicles were checked using fluorescence or confocal microscopy. Furthermore, hormone levels of estrogen (pmol/L) and progesterone (nmol/L) were determined.

Results: When comparing the SR of follicles among the four groups, it was observed that on day 6, the study groups utilizing the polycaprolactone scaffold with bpV(HOpic) in the medium (SR: 0.48 ± 0.18; p = 0.004) or functionalized in the scaffold (SR: 0.50 ± 0.20; p = 0.003) exhibited significantly higher survival rates compared to the group using only the polyethylene terephthalate membrane (SR: 0). On day 10, a significantly higher survival rate was only noted when comparing the polycaprolactone scaffold with bpV(HOpic) in the medium to the polyethylene terephthalate membrane group (SR: 0.38 ± 0.20 versus 0; p = 0.007). Higher levels of progesterone were only significantly associated with better survival rates in the group with the polycaprolactone scaffold functionalized with bpV(HOpic) (p = 0.017).

Conclusion: This study demonstrates that three-dimensional polycaprolactone scaffolds improve the survival rates of isolated mice follicles in comparison with a conventional polyethylene terephthalate membrane. The survival rates slightly improve with added bpV(HOpic). Furthermore, higher rates of progesterone were also partly associated with improved survival.

Keywords: Artificial ovary; Ovarian follicle; bpV(HOpic).

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Fig. 1
Fig. 1
The morphology of ovarian follicles was categorized on days 2, 6, and 10 of culture using 4',6-diamidino-2-phenylindole (DAPI) staining. Day 2: Follicles exhibited a spherical shape with a complete granulosa cell layer. Day 6: Follicles formed clusters and gradually acquired an irregular shape. Day 10: Follicles remained partly viable. Scale bars represent 200 µm
Fig. 2
Fig. 2
Confocal microscopy images of the follicles on Day 2 inside the PET-Membrane (A) and Functionalized PCL-Scaffold (B). A Follicles show no contact or interaction to the mesh of the PET-Membrane. B: Shows the follicle on Day 2 deep inside the Functionalized PCL scaffold. Green points show living cells, the blue fibers represent the electrospun mesh with the embedded follicles and pink represent the SiR-F-actin staining. Scale bar 50 μm
Fig. 3
Fig. 3
The figure represents the mean survival of follicles of each group on each examination day along with the standard deviation. Significant differences in the post-hoc t tests are marked with an asterisk (p < 0.01: **)
Fig. 4
Fig. 4
The figure shows the survival probabilities of four study groups over time, based on the logistic regression models. The y-axis represents the calculated survival rate, with 1.0 indicating 100% vital follicles, and a value of 0 signifying the absence of viable follicles
Fig. 5
Fig. 5
The figure shows the survival probabilities of the three study groups over time, based on the logistic regression models and their respective progesterone levels, with estrogen fixed at its median value (80.53)
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References

    1. Donnez J, Dolmans M-M. Fertility preservation in women. Nat Rev Endocrinol. 2013;9(12):735–749. doi: 10.1038/nrendo.2013.205. - DOI - PubMed
    1. Gan H-W, Spoudeas HA. Long-term follow-up of survivors of childhood cancer (SIGN Clinical Guideline 132) Arch Dis Childhood-Educ Pract. 2014;99(4):138–143. doi: 10.1136/archdischild-2013-305452. - DOI - PMC - PubMed
    1. Brenner H. Long-term survival rates of cancer patients achieved by the end of the 20th century: a period analysis. Lancet. 2002;360(9340):1131–1135. doi: 10.1016/S0140-6736(02)11199-8. - DOI - PubMed
    1. Wallace WHB, Anderson RA, Irvine DS. Fertility preservation for young patients with cancer: who is at risk and what can be offered? Lancet Oncol. 2005;6(4):209–218. doi: 10.1016/S1470-2045(05)70092-9. - DOI - PubMed
    1. Dittrich R, Kliesch S, Schüring A, Balcerek M, Baston-Büst DM, Beck R, Beckmann MW, et al. Fertility preservation for patients with malignant disease. Guideline of the DGGG, DGU and DGRM (S2k-Level, AWMF Registry No 015/082, November 217)–Recommendations and statements for girls and women. Geburtshilfe Frauenheilkd. 2018;78(06):567–584. doi: 10.1055/a-0611-5549. - DOI - PMC - PubMed