Investigating and Modelling an Engineered Millifluidic In Vitro Oocyte Maturation System Reproducing the Physiological Ovary Environment in the Sheep Model

Cells. 2022 Nov 15;11(22):3611. doi: 10.3390/cells11223611.

Abstract

In conventional assisted reproductive technologies (ARTs), oocytes are in vitro cultured in static conditions. Instead, dynamic systems could better mimic the physiological in vivo environment. In this study, a millifluidic in vitro oocyte maturation (mIVM) system, in a transparent bioreactor integrated with 3D printed supports, was investigated and modeled thanks to computational fluid dynamic (CFD) and oxygen convection-reaction-diffusion (CRD) models. Cumulus-oocyte complexes (COCs) from slaughtered lambs were cultured for 24 h under static (controls) or dynamic IVM in absence (native) or presence of 3D-printed devices with different shapes and assembly modes, with/without alginate filling. Nuclear chromatin configuration, mitochondria distribution patterns, and activity of in vitro matured oocytes were assessed. The native dynamic mIVM significantly reduced the maturation rate compared to the static group (p < 0.001) and metaphase II (MII) oocytes showed impaired mitochondria distribution (p < 0.05) and activity (p < 0.001). When COCs were included in a combination of concave+ring support, particularly with alginate filling, oocyte maturation and mitochondria pattern were preserved, and bioenergetic/oxidative status was improved (p < 0.05) compared to controls. Results were supported by computational models demonstrating that, in mIVM in biocompatible inserts, COCs were protected from shear stresses while ensuring physiological oxygen diffusion replicating the one occurring in vivo from capillaries.

Keywords: computational models; in vitro maturation; intracellular reactive oxygen species; millifluidic culture; mitochondria; oocyte; ovary environment; oxygen diffusion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alginates / pharmacology
  • Animals
  • Female
  • In Vitro Oocyte Maturation Techniques* / methods
  • Oocytes / physiology
  • Ovary*
  • Oxygen
  • Sheep

Substances

  • Oxygen
  • Alginates

Grants and funding

This research was funded by: Research Grant n. VERI00086 in the framework of the Project “Demetra” (Dipartimenti di Eccellenza 2018–2022, CUP code: C46C18000530001), funded by the Italian Ministry for Education, University, and Research; Unione europea—FSE-REACT-EU, PON Ricerca e Innovazione 2014-2020 DM1062/2021; UE—FSE-FSER, PON RI 2014-2020 Action I.1—“Innovative doctoral of industrial interest”—a.a. 2016/2017, XXXII cycle, PhD Program “Functional and Applied Genomics and Proteomics (DOT1302781”—Grant n°2).