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Effect of Previous Chemotherapy on the Quality of Cryopreserved Human Ovarian Tissue In Vitro.

Asadi Azarbaijani B, Sheikhi M, Oskam IC, Nurmio M, Laine T, Tinkanen H, Mäkinen S, Tanbo TG, Hovatta O, Jahnukainen K - PLoS ONE (2015)

Bottom Line: Increasing dose of alkylating agents was identified by multivariate linear regression analysis as an independent predictor of a higher density of atretic follicles, whereas increasing age of the patient predicted a better outcome with less follicle atresia and a higher density of maturing follicles.The results indicate that fertility preservation should be carried out, if possible, before initiation of alkylating agents in order to guarantee better in vitro survival of ovarian follicles.In addition, ovarian samples from younger girls show lower viability and fewer developing follicles in culture.

View Article: PubMed Central - PubMed

Affiliation: Women and Children's Division, Oslo University Hospital, Rikshospitalet, Oslo, Norway; University of Oslo, Oslo, Norway.

ABSTRACT

Background: Cryopreservation of ovarian tissue has been widely accepted as an option for fertility preservation among cancer patients. Some patients are exposed to chemotherapy prior to ovarian tissue cryopreservation. Consequently, assessment of the developmental capacity of human ovarian tissue after chemotherapy is of primary importance.

Materials: In order to study the impact of previous chemotherapy on in vitro development and viability of ovarian follicles, quality control samples from 34 female cancer patients at median age of 15 years (range 1‒35), cryopreserved for fertility preservation before (n = 14) or after (n = 20) initiation of chemotherapy, were thawed and cultured for 7 days. The morphology and developmental stages of ovarian follicles were studied by light microscopy before and after culture. Possible associations between follicular densities, age and exposure to alkylating agents, expressed as cyclophosphamide equivalent dose (CED) were tested.

Results: Exposure to chemotherapy significantly impaired the survival and development of ovarian follicles in culture. After seven days, significantly higher densities of intermediary, primary and secondary follicles and lower densities of atretic follicles was detected in the samples collected before chemotherapy. Increasing dose of alkylating agents was identified by multivariate linear regression analysis as an independent predictor of a higher density of atretic follicles, whereas increasing age of the patient predicted a better outcome with less follicle atresia and a higher density of maturing follicles.

Conclusion: This study provides quantitative in vitro evidence of the impact of chemotherapy on developmental capacity of cryopreserved human ovarian tissue. The results indicate that fertility preservation should be carried out, if possible, before initiation of alkylating agents in order to guarantee better in vitro survival of ovarian follicles. In addition, ovarian samples from younger girls show lower viability and fewer developing follicles in culture.

No MeSH data available.


Related in: MedlinePlus

Representative images of ovarian cortex before and after seven days of culture (magnification ×40), from a 15-year-old girl with lymphoma and no chemotherapy (A,B), and from a 2-year-old girl with neuroblastoma exposed to CED of 7200 mg/m2 (C,D).a) Intact primordial follicle, b) intact secondary follicle, c) influenced primordial follicle, d) atretic follicle.
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pone.0133985.g001: Representative images of ovarian cortex before and after seven days of culture (magnification ×40), from a 15-year-old girl with lymphoma and no chemotherapy (A,B), and from a 2-year-old girl with neuroblastoma exposed to CED of 7200 mg/m2 (C,D).a) Intact primordial follicle, b) intact secondary follicle, c) influenced primordial follicle, d) atretic follicle.

Mentions: Follicles were classified as primordial, intermediary, primary and secondary (Fig 1) [26;27]. They were further classified as intact, influenced and atretic in order to evaluate the quality of the ovarian cortical pieces (Fig 1). Intact follicles were defined as those with an intact basement membrane attached to granulosa cells and without contraction of the cytoplasm or any pyknotic nuclei. The oocytes were in close contact with the surrounding granulosa cells, and the granulosa cells were without pyknotic nuclei or any signs of shrinkage or swelling. Influenced follicles were defined as those having intact nuclei and membranes of the oocyte, with less than 50% detachment of the oocyte from surrounding granulosa cells and/or less than 10% vacuolization in cytoplasm, and less than 50% atretic granulosa cells. The follicles were defined as atretic if the nucleus or more than 50% of any of the follicle structures described above were pyknotic. All intact and influenced primordial, intermediary, primary and secondary follicles were evaluated separately in each category.


Effect of Previous Chemotherapy on the Quality of Cryopreserved Human Ovarian Tissue In Vitro.

Asadi Azarbaijani B, Sheikhi M, Oskam IC, Nurmio M, Laine T, Tinkanen H, Mäkinen S, Tanbo TG, Hovatta O, Jahnukainen K - PLoS ONE (2015)

Representative images of ovarian cortex before and after seven days of culture (magnification ×40), from a 15-year-old girl with lymphoma and no chemotherapy (A,B), and from a 2-year-old girl with neuroblastoma exposed to CED of 7200 mg/m2 (C,D).a) Intact primordial follicle, b) intact secondary follicle, c) influenced primordial follicle, d) atretic follicle.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520548&req=5

pone.0133985.g001: Representative images of ovarian cortex before and after seven days of culture (magnification ×40), from a 15-year-old girl with lymphoma and no chemotherapy (A,B), and from a 2-year-old girl with neuroblastoma exposed to CED of 7200 mg/m2 (C,D).a) Intact primordial follicle, b) intact secondary follicle, c) influenced primordial follicle, d) atretic follicle.
Mentions: Follicles were classified as primordial, intermediary, primary and secondary (Fig 1) [26;27]. They were further classified as intact, influenced and atretic in order to evaluate the quality of the ovarian cortical pieces (Fig 1). Intact follicles were defined as those with an intact basement membrane attached to granulosa cells and without contraction of the cytoplasm or any pyknotic nuclei. The oocytes were in close contact with the surrounding granulosa cells, and the granulosa cells were without pyknotic nuclei or any signs of shrinkage or swelling. Influenced follicles were defined as those having intact nuclei and membranes of the oocyte, with less than 50% detachment of the oocyte from surrounding granulosa cells and/or less than 10% vacuolization in cytoplasm, and less than 50% atretic granulosa cells. The follicles were defined as atretic if the nucleus or more than 50% of any of the follicle structures described above were pyknotic. All intact and influenced primordial, intermediary, primary and secondary follicles were evaluated separately in each category.

Bottom Line: Increasing dose of alkylating agents was identified by multivariate linear regression analysis as an independent predictor of a higher density of atretic follicles, whereas increasing age of the patient predicted a better outcome with less follicle atresia and a higher density of maturing follicles.The results indicate that fertility preservation should be carried out, if possible, before initiation of alkylating agents in order to guarantee better in vitro survival of ovarian follicles.In addition, ovarian samples from younger girls show lower viability and fewer developing follicles in culture.

View Article: PubMed Central - PubMed

Affiliation: Women and Children's Division, Oslo University Hospital, Rikshospitalet, Oslo, Norway; University of Oslo, Oslo, Norway.

ABSTRACT

Background: Cryopreservation of ovarian tissue has been widely accepted as an option for fertility preservation among cancer patients. Some patients are exposed to chemotherapy prior to ovarian tissue cryopreservation. Consequently, assessment of the developmental capacity of human ovarian tissue after chemotherapy is of primary importance.

Materials: In order to study the impact of previous chemotherapy on in vitro development and viability of ovarian follicles, quality control samples from 34 female cancer patients at median age of 15 years (range 1‒35), cryopreserved for fertility preservation before (n = 14) or after (n = 20) initiation of chemotherapy, were thawed and cultured for 7 days. The morphology and developmental stages of ovarian follicles were studied by light microscopy before and after culture. Possible associations between follicular densities, age and exposure to alkylating agents, expressed as cyclophosphamide equivalent dose (CED) were tested.

Results: Exposure to chemotherapy significantly impaired the survival and development of ovarian follicles in culture. After seven days, significantly higher densities of intermediary, primary and secondary follicles and lower densities of atretic follicles was detected in the samples collected before chemotherapy. Increasing dose of alkylating agents was identified by multivariate linear regression analysis as an independent predictor of a higher density of atretic follicles, whereas increasing age of the patient predicted a better outcome with less follicle atresia and a higher density of maturing follicles.

Conclusion: This study provides quantitative in vitro evidence of the impact of chemotherapy on developmental capacity of cryopreserved human ovarian tissue. The results indicate that fertility preservation should be carried out, if possible, before initiation of alkylating agents in order to guarantee better in vitro survival of ovarian follicles. In addition, ovarian samples from younger girls show lower viability and fewer developing follicles in culture.

No MeSH data available.


Related in: MedlinePlus