Short-term exposure of human ovarian follicles to cyclophosphamide metabolites seems to promote follicular activation in vitro.
How chemotherapy affects dormant ovarian primordial follicles is unclear. The 'burnout' theory, studied only in mice, suggests cyclophosphamide enhances primordial follicle activation. Using 4-hydroperoxycyclophosphamide (4hc) and phosphoramide mustard (PM), this study assessed how the active cyclophosphamide metabolites 4-hydroxycyclophosphamide (4-OHC) and PM, affect human primordial follicles. Frozen-thawed human ovarian samples were sliced and cultured with basic culture medium (cultured controls) or with 4hc/PM (3 µmol/l/10 µmol/l) (treated samples) for 24-48 h. Follicular counts and classification, Ki67 and anti-Müllerian hormone (AMH) immunohistochemistry and an apoptosis assay were used for evaluation, and 17β-oestradiol and AMH were measured in spent media samples. Generally, there was primordial follicle decrease and elevated developing follicle rates in treated samples compared with cultured (P = 0.04 to P < 0.0005) and uncultured controls (P < 0.05 to P < 0.0001). No traces of apoptosis were found. There were almost twicethe levels of AMH and 17β-oestradiol in treated compared with untreated samples (AMH with 4hc 3 µmol/l; P = 0.04). All follicles stained positively for AMHincluded treated samples. Ki67 positive staining was noted in all samples. Cyclophosphamide metabolites seem to enhance human primordial follicle activation to developing follicles, in vitro. Study findings support the 'burnout' theory as the mechanism of chemotherapy-induced ovarian toxicity.
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel.
Yechezkel
Y
Lande
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel.
Benjamin
B
Fisch
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel; Department of Obstetrics and Gynecology, The Chaim Sheba Medical, Ramat Gan, and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Abraham
A
Tsur
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel; IVF Unit, Department of Obstetrics and Gynecology, Edith Wolfson Medical Center, Holon, and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Jacob
J
Farhi
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel.
Roni
R
Prag-Rosenberg
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel.
Avi
A
Ben-Haroush
The Felsenstein Medical Research Center, Sackler Faculty of Medicine, Tel Aviv University, Israel.
Gania
G
Kessler-Icekson
The Felsenstein Medical Research Center, Sackler Faculty of Medicine, Tel Aviv University, Israel.
Muayad A
MA
Zahalka
Department of Basic and Social Sciences, Albany College of Pharmacy and Health Sciences, Albany, NY, USA.
Susan M
SM
Ludeman
Infertility and IVF Unit, Beilinson's Women Hospital, Rabin Medical Center Petach Tikva and Sackler Faculty of Medicine, Tel Aviv University, Israel. Electronic address: ronita@clalit.org.il.
Ronit
R
Abir
D000293:Adolescent; D054304:Anti-Mullerian Hormone / Q000627:therapeutic use; D002648:Child; D015925:Cryopreservation; D003470:Culture Media; D003520:Cyclophosphamide / Q000031:analogs & derivatives / Q000627:therapeutic use*; D046149:Embryo Culture Techniques; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005615:Freezing; D006801:Humans; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D019394:Ki-67 Antigen / Q000378:metabolism; D006080:Ovarian Follicle / Q000187:drug effects*; D010053:Ovary / Q000378:metabolism; D010752:Phosphoramide Mustards / Q000627:therapeutic use; D013997:Time Factors
D003470:Culture Media; D007166:Immunosuppressive Agents; D019394:Ki-67 Antigen; D010752:Phosphoramide Mustards; C030090:phosphoramide mustard; C012358:4-hydroxycyclophosphamide; D004958:Estradiol; D054304:Anti-Mullerian Hormone; D003520:Cyclophosphamide
4-hydroperoxycyclophamide (4hc); 4-hydroxycyclophosphamide(4-OHC); cyclophosphamide metabolites; developing follicles; human primordial follicles; in vitro; phosphoramide mustard (PM); the ‘burnout’ theory