In vitro mouse preantral follicle development in 2D and suspension culture: α-MEM vs SAGE 1-step

Area of Science:

• Reproductive Biology
• Assisted Reproductive Technology (ART)
• Cell Culture Systems

Background:

• In vitro follicle culture is crucial for fertility preservation but remains challenging.
• Developing healthier, viable eggs outside the body is a key goal for ART.

Purpose of the Study:

• To evaluate the impact of α-MEM and SAGE 1-Step media on mouse preantral follicle development.
• To compare two-dimensional (2D) and suspension culture systems for oocyte maturation.
• To optimize conditions for producing high-quality oocytes for fertility preservation.

Main Methods:

• Mouse preantral follicles were cultured for 13 days in α-MEM or SAGE 1-Step using 2D or suspension systems.
• Oocyte maturation was induced with hCG.
• Follicular growth, antrum formation, meiotic spindle organization, and gene expression (maturation, cumulus expansion, apoptosis) were assessed.

Main Results:

• SAGE 1-Step medium significantly improved oocyte maturation in both 2D and suspension cultures.
• Suspension culture significantly increased normal meiotic spindle organization and reduced apoptosis (decreased Bax/Bcl2 ratio).
• Suspension culture upregulated Adamts1 and downregulated Bax gene expression.

Conclusions:

• SAGE 1-Step medium enhances oocyte maturation.
• Suspension culture preserves meiotic spindle architecture and reduces apoptosis.
• Combining SAGE 1-Step medium with suspension culture optimizes the microenvironment for producing high-quality oocytes.