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Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System
Mae W Healy1,2, Shelley N Dolitsky1, Maria Villancio-Wolter3
1Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
Micromachines
|April 3, 2021
Summary
Researchers developed a novel microfluidic system to create 3D ovarian follicles, successfully encapsulating oocytes and supporting their environment for potential fertility applications.
Area of Science:
- Reproductive Biology
- Biomaterials Engineering
- Microfluidics
Background:
- Mammalian oocyte development requires a specific microenvironment provided by granulosa and theca cells within ovarian follicles.
- Current methods for studying oocytes often fail to fully replicate this complex in vivo environment.
- 3D culture systems offer a promising approach to better mimic native ovarian structures.
Purpose of the Study:
- To engineer a 3D ovarian follicle model using a microfluidic system.
- To encapsulate primary and early secondary oocytes within a biomimetic granulosa and theca cell environment.
- To assess the viability, structural integrity, and endocrine function of the engineered follicles.
Main Methods:
- Fabrication of 3D two-layer capsules using a microfluidic system with controlled flow rates.
- Encapsulation of murine granulosa cells in a collagen-alginate core and murine theca cells in an alginate shell.
- Assessment of cell viability, hormonal production (estradiol, progesterone, androstenedione), and oocyte integrity over 27 days.
- Confocal microscopy for verifying cell compartmentalization.
Main Results:
- Successful creation of 3D capsules with distinct core (granulosa) and shell (theca) layers, confirmed by confocal microscopy.
- Sustained cell viability greater than 78% for up to 27 days within the capsules.
- Demonstrated endocrine function through the production of estradiol, progesterone, and androstenedione.
- Successful encapsulation of primary and early secondary oocytes, maintaining their size and cellular structure.
Conclusions:
- The microfluidic system effectively recapitulates the essential two-compartment structure of mammalian ovarian follicles.
- The engineered 3D ovarian follicles provide a supportive microenvironment for oocyte development and exhibit sustained endocrine function.
- This novel system is adaptable for sterile, high-throughput applications in reproductive research and potentially in vitro fertilization.
