Mouse in vitro follicle culture bioassay for fundamental and translational research on oocyte developmental capacity
Scope of the method
- Human health
- Basic Research
- Translational - Applied Research
- In vitro - Ex vivo
- Animal derived cells / tissues / organs
Description
- in vitro
- oogenesis
- folliculogenesis
- oocyte maturation
- follicle culture
- fertility preservation
- in vitro oocyte maturation
- assisted reproductive technology
- oocyte quality
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.
- - 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).
- History of use
- Internally validated
- Published in peer reviewed journal
Pros, cons & Future potential
- - 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.
Developmental capacity of the cultured oocytes is still inferior compared to in vivo developed counterparts. The system would benefit from further optimization.
- 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
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
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
Contact person
Ellen AnckaertOrganisations
Vrije Universiteit Brussel (VUB)Pathology/molecular and cellular medicine
Belgium
