Effect of cryopreservation and transplantation on the expression of kit ligand and anti-Mullerian hormone in human ovarian tissue.
BACKGROUND
Although cryopreservation and transplantation of ovarian tissue represent a promising alternative to safeguard fertility in cancer patients, low recovery rates of oocytes aspirated from antral follicles and a significant number of empty follicles have been observed in women with transplanted frozen-thawed ovarian tissue. In order to understand how freezing and/or grafting may affect follicular development, the follicular expression of kit ligand (KL) and anti-Müllerian hormone (AMH), two key factors activating and inhibiting follicle growth, were assessed after long-term grafting in severe combined immunodeficient (SCID) mice.


METHODS
Ovarian biopsies from eight patients were used for fresh and frozen-thawed tissue xenografting in 13 SCID mice for a period of 28 weeks, including 2 weeks of gonadotrophin stimulation. KL, AMH and proliferating cell nuclear antigen (PCNA) immunostaining were quantified before and after grafting in the two treatment groups (fresh and frozen-thawed grafted ovarian tissue).


RESULTS
Lower expression of KL was found in primordial and primary follicles after grafting of both fresh and frozen-thawed tissue. Consistent expression of AMH was found in most growing follicles at a similar rate in both graft types. In fresh and frozen-thawed grafts, 13-14% of primordial follicles were PCNA-positive, indicating a similar maintenance of quiescent follicles despite follicle activation.


CONCLUSIONS
Grafting and/or gonadotrophin stimulation appear to affect the follicular expression of KL, which may alter oocyte quality. AMH expression in growing follicles after ovarian tissue transplantation may be one of the factors contributing to the preservation of resting follicles in 28-week-old grafts.
Department of Gynecology, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Avenue Emmanuel Mounier 52, 1200 Brussels, Belgium.
Anu
A
David
Anne
A
Van Langendonckt
Sébastien
S
Gilliaux
Marie-Madeleine
MM
Dolmans
Jacques
J
Donnez
Christiani A
CA
Amorim
D000818:Animals; D054304:Anti-Mullerian Hormone / Q000378:metabolism*; D015925:Cryopreservation; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006062:Gonadotropins / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D010053:Ovary / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology / Q000637:transplantation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D019089:Stem Cell Factor / Q000378:metabolism*; D014183:Transplantation, Heterologous / Q000473:pathology
D006062:Gonadotropins; D018809:Proliferating Cell Nuclear Antigen; D019089:Stem Cell Factor; D054304:Anti-Mullerian Hormone