title: The impact of culture conditions on early follicle recruitment and growth from human ovarian cortex biopsies in vitro.
abstract: OBJECTIVE
  To investigate the effects of a dynamic fluidic culture system on early in vitro folliculogenesis in standardized ovarian cortex biopsies.
  
  
  METHODS
  Cortical small strips were cultured for 6 days in a conventional static or in a dynamic fluidic culture system.
  
  
  METHODS
  University-affiliated laboratory with an associated cryobank facility.
  
  
  METHODS
  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.
  
  
  METHODS
  None.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Gynecologic Endocrinology and Reproductive Medicine, University Women's Hospital, Bonn, Germany. jana.liebenthron@ukb.uni-bonn.de
authors.forename: Jana
authors.initials: J
authors.lastname: Liebenthron
authors.forename: Maria
authors.initials: M
authors.lastname: Köster
authors.forename: Christina
authors.initials: C
authors.lastname: Drengner
authors.forename: Jochen
authors.initials: J
authors.lastname: Reinsberg
authors.forename: Hans
authors.initials: H
authors.lastname: van der Ven
authors.forename: Markus
authors.initials: M
authors.lastname: Montag
mesh_terms: D000328:Adult; D001706:Biopsy; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D005260:Female; D059247:Fertility Preservation; D006801:Humans; D059471:In Vitro Oocyte Maturation Techniques; D008954:Models, Biological; D009865:Oocytes; D006080:Ovarian Follicle / Q000166:cytology*; D010053:Ovary / Q000166:cytology*; D046509:Tissue Culture Techniques; D059467:Transcriptome
keywords: Ovarian cortical tissue; early folliculogenesis; fertility preservation; in vitro fluid culture; oncofertility