The impact of culture conditions on early follicle recruitment and growth from human ovarian cortex biopsies in vitro

The use of a dynamic fluidic culture system for standardized ovarian cortex biopsies enables both follicle survival and initiation of the earliest follicle development to be better achieved.


Jana Liebenthron, M.Sc., Maria Köster, D.V.M., Christina Drengner, Jochen Reinsberg, Ph.D., Hans van der Ven, M.D., Markus Montag, Ph.D.

Volume 100, Issue 2, Pages 483-491.e5, August 2013



To investigate the effects of a dynamic fluidic culture system on early in vitro folliculogenesis in standardized ovarian cortex biopsies.


Cortical small strips were cultured for 6 days in a conventional static or in a dynamic fluidic culture system.


University-affiliated laboratory with an associated cryobank facility.


Ovarian cortex from postpuberal female cancer patients (26.1 ± 1.3 y) who opted for cryopreservation of their tissue for fertility protection before gonadotoxic cancer therapy. With informed consent of the Institutional Ethics Committee, part of the tissue was available for patient-related research studies.



Main Outcome Measure(s):

The viability and proliferative capacity of the cortex biopsies were evaluated by chemiluminescent microparticle immunoassay for detection of in vitro produced E2 and P in the supernate, by viable follicle counting via calcein staining, by histologic analyses, and by total RNA preparation and reverse transcription for real-time polymerase chain reaction of selected early folliculogenesis genes.


The data support the notion that early follicle development can be better achieved in vitro in a dynamic fluidic culture system. The findings are based on the presence of more viable follicles, higher expression levels of early folliculogenesis genes KIT-L, INHB, and GDF9, and the absence of premature luteinization of follicles.


This study provides evidence that dynamic fluidic culture is a promising approach for investigating early follicular recruitment and growth in cortical biopsies. It may serve as a first step in a multistep culture system to design a complex in vitro system for complete folliculogenesis.

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