Germ cells markers were detected in premature ovarian failure modeling in mice: hope to fertility

Fatemeh Abedi,1 Rouhollah fathi,2,* Naeimeh abtahi,3 Farideh eivazhkani,4 Elham abedheydari,5 Khadijeh bahrebar,6

1. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
2. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
3. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
4. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
5. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
6. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.

Abstract


Introduction

Ovary is one of the endocrine organs of female reproductive system which produces necessary steroids and peptide hormones for puberty and the menstrual cycle. chemo/radiotherapy leads to premature ovarian failure (pof) due to reducing follicles, ovulation and fertility. cyclophosphamide used in the treatment of cancer is one of the usual chemotherapy and alkylating agents which causes dna break in different types of the cells. this study focused on cyclophosphamide and busulfan effects on mouse ovary to produce pof model for the next studies on fertility preservation.

Methods

Nmri female mice were treated with daily intraperitoneal injection of 75 mg/kg (group 1) and 100 mg/kg (group 2) cyclophosphamide,100 mg/kg cyclophosphamide first day, 20 mg/kg daily for 13 days (group3), 100mg/kg cyclophosphamide first day, 50 mg/kg busulfan daily for 13 days (group 4), 75 mg/kg cyclophosphamide and 30 mg/kg busulfan daily for 14 days (group 5) kept in separate specific cages for each group for 14 days and control group which received no chemotherapy drugs. change body weight, smear test and histology assessment were evaluated for all experimental and control groups before and after chemotherapy. estradiol(e2), follicle stimulating hormone(fsh) and real time pcr for oct4, blimp1, dazl and gdf9 for the best group of pof model were evaluated.

Results

The percentage of vital primordial follicles in all experimental groups was significantly lower than control group (p <0.001). in the second experimental group, the percentage of vital follicles in different stages of primordial, pre antral and antral was significantly decreased (p <0.01) compared to the control group. alterations of body weight demonstrated notable decrease in groups 1and 2 as compared to control group (p<0.05). the irregularity of the estrous cycle, remarkable increase of fsh and significant reduce of estradiol secretion in the second group compared to the control group was shown (p <0.05), which confirmed the creation of the pof model successfully in second group. also, the result of real time pcr showed significant increase of oct-4 and dazl in the second group compared to control group (p <0.05).

Conclusion

According to these results, treatment of 100 mg/kg cyclophosphamide for 14 days caused a powerful damage on ovarian activity and reduce follicular reservation as compared to all of experimental groups. further, lack of corpus luteum in second group showed folliculogenesis disrupt and approved creation of mouse premature ovarian failure model. this study showed, in regard to damage of ovarian tissue by chemotherapy drugs, there are still germ cells in the pof model and showed a hopeful and certain future for treating infertility in cancer patients after chemotherapy.

Keywords

Premature ovarian failure, ovarian reserve, folliculogenesis, chemotherapy.