Ovarian puncture alters genes but not follicle growth in cyclophosphamide mice model

Cyclophosphamide administration often causes infertility. Ovarian puncture (OP), a component of in vitro fertilization, is performed to obtain oocytes. Our previous study showed that OP stimulated follicular activation in mice. However, its effects on folliculogenesis in cyclophosphamide-treated animals remain unclear. This study aimed to evaluate the impact of OP on follicular growth and gene expression. Female C57BL/06J (n=52) were divided into groups (control, cyclo, and cyclo+Pc). Cyclophosphamide was administered in four doses into the cyclo and cyclo+Pc groups (150 mg/kg), and OP was performed three days after the last dose and 96 h before euthanasia in the cyclo+Pc group. There was no difference in body weight; however, ovarian weight was higher in cyclo+Pc compared to cyclo. The number of all follicle classes was similar between cyclo+Pc and cyclo. Caspase-3-positive granulosa cells (%) in antral follicles were increased in cyclo compared to control and reduced in cyclo+Pc compared to control. Amh expression was increased in the cyclo+Pc compared with the cyclo. Pten expression was reduced in cyclo+Pc, whereas Kitl expression was increased. Bax expression was reduced, and there was no change in Casp3 expression due to OP. Moreover, collagen deposition was increased in Cyclo+Pc compared with the control and cyclo groups. However, the treatments did not change the ovulation response, oocytes, or follicular morphometrics. Our experimental study demonstrates, for the first time, that OP is a safe procedure that can be performed in cyclophosphamide treatment in mice. Lay summary Infertility is a concern for cancer patients who wish to conceive after completing chemotherapy treatment. Puncturing the ovary is part of the IVF procedure, aimed at collecting eggs to fertilize and transferring them to the uterus. We decided to study whether the inflammation and tissue damage caused by puncturing the ovaries could stimulate the collection of healthy eggs in patients who have gone through chemotherapy. To this end, we used female mice that were treated with a frequently used treatment for breast cancer, along with ovarian puncture. We discovered the alterations did not affect follicle growth or egg retrieval. With these findings, we demonstrated, for the first time, the ability of ovarian puncture to bring about healthy egg retrieval, improving quality of life after cancer.