International Conference onStem Cells, Regenerative Medicine and Tissue Engineering

Title: Insight on AEC and AFMSC mechanisms Preserving male fertility after induced Varicocele in Rat experimental model

Abstract:

Statement: Amniotic membrane and amniotic fluid-derived cells are considered a promising source of stem cells for the development of regenerative medicine techniques (1-4), yet there is no evidence that they may treat male infertility illnesses like varicocele (VAR), one of the main causes of male infertility which accounts for about 40% of primary and 80% of secondary male factor infertility, respectively (5). Despite the encouraging paracrine, anti-inflammatory, and immune regulatory therapeutic role of this stem cell source (1-4,6,7), no evidence on male fertility has been collected to date.
Aim: The current study aimed to examine the effects of two distinct cell sources, human Amniotic Fluid Mesenchymal Stromal Cells (hAFMSCs) and amniotic epithelial cells (hAECs), on male fertility outcomes in a validated rat-induced VAR model (5) selected for its high translational value due to its capability of replicating several aspects of the human pathology, including alterations in testicular blood flow, spermatogenesis, and immune response.
Methodology & Experimental plan: hAECs and hAFMSCs were marked with PKH26 vital cell membrane dye for in vivo cell tracking and then transplanted (0.5x106 in 50 ul of vehicle/each type of cells) in left testis with intratesticular injection in VAR rats groups: +hAEC (n=10) and +hAFMSC (n=10) respectively. Healthy (CTR, n=10) and VAR rats (+vehicle alone) groups were considered. The influence of cell transplantation was first assessed considering the long-term impact on rat fertility by recording the newborn number after two sequential matings carried out 120 days after surgical procedures. Then, the mechanisms related to fertility outcomes were in-depth analyzed by focusing the attention on testicular morphology, endocannabinoid system (ECS) expression profile, and inflammatory response, in parallel, with the evaluation of homing of hAFMSCs and hAECs and testis recovery upon their transplantation.
Results: hAECs and hAFMSC, were able to recover fertility in a VAR experimental rat model, even if depending on the type of stem cell used. Both cell types survived 120 days post-transplantation by modulating the ECS main components, (NAPE-PLD, DAGL , and MAGL; p< 0.01 vs. both CTR and +vehicle) as well as the intracellular TRPV1 receptor (+hAECs, p < 0.001 vs. +vehicle), promoting pro-regenerative CD206 positive M2 macrophages (Mφ) recruitment and a favorable anti-inflammatory IL10 expression pattern (p< 0.001 hAECs and hAFMSCs vs. +vehicle). Of note, hAECs resulted in being more effective in restoring rat fertility rate by enhancing both structural and immune response mechanisms. Moreover, immunofluorescence analysis revealed that hAECs contributed to CYP11A1 expression after transplantation, whereas hAFMSCs moved towards the expression of Sertoli cell marker, SOX9, confirming a different contribution into the mechanisms leading to testis homeostasis (8).
Conclusion & Significance: These findings highlight, for the first time, a distinct role of amniotic membrane and amniotic fluid-derived cells in male reproduction, thus proposing innovative targeted stem-based regenerative medicine protocols for remedying high-prevalence male infertility conditions such as VAR.

Biography:

Annunziata Mauro is an Associate Professor at the University of Teramo. She is the coordinator of the Bachelor's degree in Biotechnology. Her research interests mainly focus on studying stem cells of amniotic origin and their application in regenerative medicine and tissue engineering in species of veterinary interest with translational potential in humans. She translates her intellectual and technical knowledge developed in the fields of Cell Biology and Reproductive Biology into the study of amniotic stem cells and their therapeutic potential role, applying the latest and most innovative research techniques and operating within national and international research networks. She participates at National (PRIN, FFABR, PNRR) and European (H2020-EU.1.3.1.-MSCA-ITN 2015-EJD: REP-BIOTECH; H2020-EU.1.3.4.- MSCA-COFUND-2015-DP: REP-EAT; H2020-MSCA-ITN-2020-MSCA-ITN EJD: P4FIT) research projects. She has built her university commitment after years of experience in research, evaluation, teaching, and administration in academic institutions.