title: Healthy early preantral follicle can be obtained in a culture of frozen-thawed human ovarian tissue of 32 weeks.
abstract: The objective of this study was to report morphological and functional evidence of a well-preserved preantral follicle recovered from human frozen-thawed ovarian tissue in a long-term culture. The tissue was originally obtained from a 26-year-old woman with breast cancer. The ovarian cortex was collected by laparoscopy and frozen/thawed and cultured for 32 weeks in minimum essential medium alpha-MEM, supplemented with insulin transferrine selenite (ITS), human serum (HS), antibiotics, follicle-stimulating hormone (FSH). and N-acetyl cysteine (NAC). Thawed tissue samples were examined by light microscopy (LM), transmission electron microscopy (TEM), and real-time RT-PCR. LM examination of cortical pieces after 32 weeks of culture showed a healthy early preantral follicle; TEM and real-time PCR confirmed its good state of preservation. The synergy in action of NAC and FSH plays an important role in follicle growth of ovarian tissue cultures. For the first time a well-preserved preantral follicle was found in a culture of frozen-thawed human ovarian tissue.
authors.affiliation: Human Reproduction Medicine Unit, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy. raffaella.fabbri@unibo.it
authors.forename: Raffaella
authors.initials: R
authors.lastname: Fabbri
authors.forename: Gianandrea
authors.initials: G
authors.lastname: Pasquinelli
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Montanaro
authors.forename: Bruno
authors.initials: B
authors.lastname: Mozzanega
authors.forename: Valentina
authors.initials: V
authors.lastname: Magnani
authors.forename: Federica
authors.initials: F
authors.lastname: Tamburini
authors.forename: Stefano
authors.initials: S
authors.lastname: Venturoli
authors.forename: Declan
authors.initials: D
authors.lastname: Keane
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000328:Adult; D055430:Bone Morphogenetic Protein 15; D018929:Cell Culture Techniques / Q000379:methods*; D015925:Cryopreservation; D003470:Culture Media / Q000737:chemistry; D005260:Female; D055429:Growth Differentiation Factor 9; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis; D046529:Microscopy, Electron, Transmission; D006080:Ovarian Follicle / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure*; D010053:Ovary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors