Three-Dimensionally Printed Agarose Micromold Supports Scaffold-Free Mouse Ex Vivo Follicle Growth, Ovulation, and Luteinization

Emily J. Zaniker; Prianka H. Hashim; Samuel Gauthier; James A. Ankrum; Hannes Campo; Francesca E. Duncan

doi:10.3390/bioengineering11070719

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Three-Dimensionally Printed Agarose Micromold Supports Scaffold-Free Mouse Ex Vivo Follicle Growth, Ovulation, and Luteinization

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Three-Dimensionally Printed Agarose Micromold Supports Scaffold-Free Mouse Ex Vivo Follicle Growth, Ovulation, and Luteinization

Emily J. Zaniker, Prianka H. Hashim, Samuel Gauthier, James A. Ankrum, Hannes Campo and Francesca E. Duncan

Bioengineering (Basel), Vol.11(7), p.719

07/15/2024

DOI: 10.3390/bioengineering11070719

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Abstract

Ex vivo follicle growth is an essential tool, enabling interrogation of folliculogenesis, ovulation, and luteinization. Though significant advancements have been made, existing follicle culture strategies can be technically challenging and laborious. In this study, we advanced the field through development of a custom agarose micromold, which enables scaffold-free follicle culture. We established an accessible and economical manufacturing method using 3D printing and silicone molding that generates biocompatible hydrogel molds without the risk of cytotoxicity from leachates. Each mold supports simultaneous culture of multiple multilayer secondary follicles in a single focal plane, allowing for constant timelapse monitoring and automated analysis. Mouse follicles cultured using this novel system exhibit significantly improved growth and ovulation outcomes with comparable survival, oocyte maturation, and hormone production profiles as established three-dimensional encapsulated in vitro follicle growth (eIVFG) systems. Additionally, follicles recapitulated aspects of in vivo ovulation physiology with respect to their architecture and spatial polarization, which has not been observed in eIVFG systems. This system offers simplicity, scalability, integration with morphokinetic analyses of follicle growth and ovulation, and compatibility with existing microphysiological platforms. This culture strategy has implications for fundamental follicle biology, fertility preservation strategies, reproductive toxicology, and contraceptive drug discovery.

Details

Title: Subtitle: Three-Dimensionally Printed Agarose Micromold Supports Scaffold-Free Mouse Ex Vivo Follicle Growth, Ovulation, and Luteinization
Creators: Emily J. Zaniker - Northwestern University
Prianka H. Hashim - Northwestern University
Samuel Gauthier - Northwestern University
James A. Ankrum - University of Iowa
Hannes Campo - Northwestern University
Francesca E. Duncan - Northwestern University
Resource Type: Journal article
Publication Details: Bioengineering (Basel), Vol.11(7), p.719
DOI: 10.3390/bioengineering11070719
ISSN: 2306-5354
eISSN: 2306-5354
Language: English
Date published: 07/15/2024
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Fraternal Order of Eagles Diabetes Research Center
Record Identifier: 9984658352402771

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