Mouse in vitro follicle culture bioassay for fundamental and translational research on oocyte developmental capacity

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Basic Research
  • Translational - Applied Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
mouse
Type of cells/tissues/organs
ovarian follicles

Description

Method keywords
  • in vitro
  • oogenesis
  • folliculogenesis
  • oocyte maturation
Scientific area keywords
  • follicle culture
  • fertility preservation
  • in vitro oocyte maturation
  • assisted reproductive technology
  • oocyte quality
Method description

Follicle Biology Laboratory has developed a well characterized and standardized MOUSE in vitro follicle culture (IFC) system. In this system, early stage ovarian follicles are cultured in vitro under physiological hormone concentrations up to fertilizable and developmentally competent mature oocytes. The follicle culture bioassay provides unique opportunities to study ovarian physiology and to assess the effects of adverse metabolic, nutritional, toxicological or environmental exposure on epigenetic reprogramming, folliculogenesis and oocyte quality. The IFC system allows identifying molecules and pathways that affect oocyte quality. Furthermore, in vitro systems for oocyte maturation (IVM) allow determining windows of sensitivity. Finally, the mouse IFC and IVM systems are a model for translational research in the context of human fertility preservation strategies and optimized IVM protocols in infertile patients.

Lab equipment
  • - Cabinet laminar flow with: hot plate and stereomicroscope equipped with a hot plate ;
  • - Inverted microscope with 40x objective and ocular scale for measurement ;
  • - Incubators (5%CO2, normal oxygen).
Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages
  • - In vitro development allows the standardized growth and maturation of high numbers of ovarian follicles at the same developmental stage. This is not achievable in vivo in an efficient way.
  • - The system has already been extensively characterized (published in peer reviewed journals).
  • - The system has great potential as a bioassay for testing exposure to adverse conditions.
Challenges

Developmental capacity of the cultured oocytes is still inferior compared to in vivo developed counterparts. The system would benefit from further optimization.

Modifications
  • We plan a project on the optimization of the IFC system.
  • Strategy:
  • - 3D culture system
  • - Incorporation of extracellular matrix components
  • - Addition of specific hormones and growth factors that might enhance developmental capacity
  • - Addition of somatic feeder cells
Future & Other applications

After optimization the IFC system will result in a bioassay with targeted endpoints for testing of culture media, pharmacological and toxicological compounds and metabolic and nutritional challenges with potential for valorization.

References, associated documents and other information

References

In vitro follicle culture in the context of IVF. Herta AC, Lolicato F, Smitz JEJ. Reproduction. 2018

Immature Oocytes from Unprimed Juvenile Mice Become a Valuable Source for Embryo Production When Using C-Type Natriuretic Peptide as Essential Component of Culture Medium. Romero S, Sánchez F, Lolicato F, Van Ranst H, Smitz J. Biol Reprod. 2016.

Culture of oocytes and risk of imprinting defects. Anckaert E, De Rycke M, Smitz J. Hum Reprod Update. 2013

Oocyte and cumulus cell transcripts from cultured mouse follicles are induced to deviate from normal in vivo conditions by combinations of insulin, follicle-stimulating hormone, and human chorionic gonadotropin. Sánchez F, Romero S, Smitz J. Biol Reprod. 2011

Mouse cumulus-oocyte complexes from in vitro-cultured preantral follicles suggest an anti-luteinizing role for the EGF cascade in the cumulus cells. Romero S, Sánchez F, Adriaenssens T, Smitz J. Biol Reprod. 2011

Associated documents

Contact person

Ellen Anckaert

Organisations

Vrije Universiteit Brussel
Follicle Biology Laboratory
Belgium
Brussels Region