Protocol for ex vivo mouse paired single-gonad screening platform for phenotypical characterization

Area of Science:

• Developmental biology
• Reproductive biology
• Organ culture

Background:

• Fetal gonad development is crucial for reproductive health.
• Short-term screening methods are needed to study gonad development.
• Existing methods may not be optimized for rapid inhibitor or modulator screening.

Purpose of the Study:

• To present a novel ex vivo paired single-gonad culture protocol.
• To optimize the protocol for short-term screening of inhibitors, metabolites, and signaling modulators.
• To provide a standardized method for reproducible gonad research.

Main Methods:

• Fetal mouse gonad dissection and paired culture setup.
• Optimization of experimental design to minimize variability.
• Detailed procedures for downstream phenotypic assessment, including immunostaining, multi-omics, and metabolic measurements.

Main Results:

• A reproducible ex vivo culture protocol for fetal mouse gonads was established.
• The protocol is suitable for short-term screening of various modulators.
• Downstream analyses allow comprehensive assessment of gonadal tissue and media.

Conclusions:

• This ex vivo paired gonad culture protocol offers a valuable tool for studying gonad development.
• The method facilitates efficient screening of compounds influencing gonadal function.
• It supports diverse downstream analyses for a holistic understanding of gonadal biology.