title: Evaluation of a human ovarian follicle isolation technique to obtain disease-free follicle suspensions before safely grafting to cancer patients.
abstract: OBJECTIVE
  To evaluate the safety of our follicle isolation procedure in a model of ovarian tissue artificially contaminated with cancer cells, then to improve the procedure to effectively eliminate malignant cells from follicle suspensions without altering viability.
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  Gynecology research unit in a university hospital.
  
  
  METHODS
  Ten women undergoing laparoscopy for benign gynecologic disease.
  
  
  METHODS
  Follicle isolation from ovarian tissue artificially contaminated with marked fluorescent leukemic cells, either by the usual pickup technique without further treatment (group 1) or by washing three times after pickup (group 2).
  
  
  METHODS
  Evidence of leukemic cells in follicle suspensions using fluorescence microscopy and quantitative real-time polymerase chain reaction, and analysis of follicle viability.
  
  
  RESULTS
  In group 1, 196 leukemic cells were detected by fluorescence microscopy out of 499 follicles retrieved, while just one leukemic cell was found among 772 follicles after three washes. The BCR-ABL fusion transcript was detected when at least 19 cells were present in follicle suspensions; four samples were positive in group 1, and all were negative in group 2. Follicle viability was similar in both groups (95.6% vs. 96.4%).
  
  
  CONCLUSIONS
  Cancer cells could inadvertently be picked up with isolated follicles in case of malignant contamination of ovarian tissue. A simple purging procedure consisting of three washes proved effective for eliminating leukemic cells while maintaining good follicle viability.
authors.affiliation: Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium; Département de Gynécologie, Cliniques Universitaires St. Luc, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Michelle
authors.initials: M
authors.lastname: Soares
authors.affiliation: Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Karima
authors.initials: K
authors.lastname: Sahrari
authors.affiliation: Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Christiani Andrade
authors.initials: CA
authors.lastname: Amorim
authors.affiliation: Départment de Biologie Clinique, Cliniques Universitaires St. Luc, Université Catholique de Louvain, Brussels, Belgium.
authors.forename: Pascale
authors.initials: P
authors.lastname: Saussoy
authors.affiliation: Société de Recherche pour l'Infertilité, Brussels, Belgium.
authors.forename: Jacques
authors.initials: J
authors.lastname: Donnez
authors.affiliation: Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium; Département de Gynécologie, Cliniques Universitaires St. Luc, Université Catholique de Louvain, Brussels, Belgium. Electronic address: marie-madeleine.dolmans@uclouvain.be.
authors.forename: Marie-Madeleine
authors.initials: MM
authors.lastname: Dolmans
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000235:genetics; D001706:Biopsy; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D005260:Female; D059247:Fertility Preservation / Q000379:methods*; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D006801:Humans; D007938:Leukemia / Q000235:genetics / Q000473:pathology*; D008856:Microscopy, Fluorescence; D006080:Ovarian Follicle / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055815:Young Adult
chemical_list: C581634:BCR-ABL1 fusion protein, human; D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D016044:Fusion Proteins, bcr-abl
keywords: Follicle isolation; leukemia; malignant cell purging; minimal disseminated disease; ovarian follicles; ovarian tissue cryopreservation