Healthy early preantral follicle can be obtained in a culture of frozen-thawed human ovarian tissue of 32 weeks.
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.
Fabbri
Raffaella
R
Human Reproduction Medicine Unit, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy. raffaella.fabbri@unibo.it
Pasquinelli
Gianandrea
G
Montanaro
Lorenzo
L
Mozzanega
Bruno
B
Magnani
Valentina
V
Tamburini
Federica
F
Venturoli
Stefano
S
Keane
Declan
D
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