Enhanced solute transport and steady mechanical stimulation in a novel dynamic perifusion bioreactor increase the efficiency of the in vitro culture of ovarian cortical tissue strips

Gionata Fragomeni; Luigi De Napoli; Vincenza De Gregorio; Vincenzo Genovese; Vincenza Barbato; Giuseppe Serratore; Giuseppe Morrone; Angela Travaglione; Andrea Candela; Roberto Gualtieri; Riccardo Talevi; Gerardo Catapano

doi:10.3389/fbioe.2024.1310696

Enhanced solute transport and steady mechanical stimulation in a novel dynamic perifusion bioreactor increase the efficiency of the in vitro culture of ovarian cortical tissue strips

Front Bioeng Biotechnol. 2024 Feb 8:12:1310696. doi: 10.3389/fbioe.2024.1310696. eCollection 2024.

Affiliations

¹ Department of Medical and Surgical Sciences, Magna Graecia University, Catanzaro, Italy.
² Department of Mechanical, Energy and Management Engineering, University of Calabria, Rende, Italy.
³ Department of Biology, University of Naples Federico II, Complesso Universitario di Monte S. Angelo, Naples, Italy.

^# Contributed equally.

Abstract

Introduction: We report the development and preliminary evaluation of a novel dynamic bioreactor to culture ovarian cortical tissue strips that leverages tissue response to enhanced oxygen transport and adequate mechanical stimulation. In vitro multistep ovarian tissue static culture followed by mature oocyte generation, fertilization, and embryo transfer promises to use the reserve of dormant follicles. Unfortunately, static in vitro culture of ovarian tissue does not promote development of primordial to secondary follicles or sustain follicle viability and thereby limits the number of obtainable mature oocytes. Enhancing oxygen transport to and exerting mechanical stimulation on ovarian tissue in a dynamic bioreactor may more closely mimic the physiological microenvironment and thus promote follicle activation, development, and viability. Materials and Methods: The most transport-effective dynamic bioreactor design was modified using 3D models of medium and oxygen transport to maximize strip perifusion and apply tissue fluid dynamic shear stresses and direct compressive strains to elicit tissue response. Prototypes of the final bioreactor design were manufactured with materials of varying cytocompatibility and assessed by testing the effect of leachables on sperm motility. Effectiveness of the bioreactor culture was characterized against static controls by culturing fresh bovine ovarian tissue strips for 7 days at 4.8 × 10^-5 m/s medium filtration flux in air at -15% maximal total compressive strain and by assessing follicle development, health, and viability. Results and Conclusions: Culture in dynamic bioreactors promoted effective oxygen transport to tissues and stimulated tissues with strains and fluid dynamic shear stresses that, although non-uniform, significantly influenced tissue metabolism. Tissue strip culture in bioreactors made of cytocompatible polypropylene preserved follicle viability and promoted follicle development better than static culture, less so in bioreactors made of cytotoxic ABS-like resin.

Keywords: bioreactor; dynamic culture; fluid dynamic stimulation; mechanical stimulation; ovarian tissue; oxygen transport.

Grants and funding

The authors declare that financial support was received for the research, authorship, and/or publication of this article. This study received funding from IVF Red srl, Via San Josemaria Escrivà 68, 81100 Caserta, Italy. Funding was also provided by Merck Healthcare KGaA, Frankfurter Str. 250, 64293 Darmstadt, Germany, within the Medical Innovation Program (MIP) agreement, and by the project CalHubRia (PSC Salute–Traiettoria 4 del Piano Operativo Salute: “Biotecnologie, Bioinformatica e Sviluppo Farmaceutico” Linea di azione 4.1 “Creazione di Hub delle Scienze della Vita”). The funders were not involved in the study design, collection, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication.