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

Capsule:
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.

Authors:
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

Abstract:

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

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

Setting:
University-affiliated laboratory with an associated cryobank facility.

Patient(s):
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.

Intervention(s):
None.

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.

Result(s):
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.

Conclusion(s):
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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