Fertility In Female Cancer Patients Research Articles

Mesenchymal stem cells promote ovarian reconstruction in mice

BackgroundStudies have shown that chemotherapy and radiotherapy can cause premature ovarian failure and loss of fertility in female cancer patients. Ovarian cortex cryopreservation is a good choice to preserve female fertility before cancer treatment. Following the remission of the disease, the thawed ovarian tissue can be transplanted back and restore fertility of the patient. However, there is a risk to reintroduce cancer cells in the body and leads to the recurrence of cancer. Given the low success rate of current in vitro culture techniques for obtaining mature oocytes from primordial follicles, an artificial ovary with primordial follicles may be a good way to solve this problem.MethodsIn the study, we established an artificial ovary model based on the participation of mesenchymal stem cells (MSCs) to evaluate the effect of MSCs on follicular development and oocyte maturation. P2.5 mouse ovaries were digested into single cell suspensions and mixed with bone marrow derived mesenchymal stem cells (BM-MSCs) at a 1:1 ratio. The reconstituted ovarian model was then generated by using phytohemagglutinin. The phenotype and mechanism studies were explored by follicle counting, immunohistochemistry, immunofluorescence, in vitro maturation (IVM), in vitro fertilization (IVF), real-time quantitative polymerase chain reaction (RT-PCR), and Terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL) assay.ResultsOur study found that the addition of BM-MSCs to the reconstituted ovary can enhance the survival of oocytes and promote the growth and development of follicles. After transplanting the reconstituted ovaries under kidney capsules of the recipient mice, we observed normal folliculogenesis and oocyte maturation. Interestingly, we found that BM-MSCs did not contribute to the formation of follicles in ovarian aggregation, nor did they undergo proliferation during follicle growth. Instead, the cells were found to be located around growing follicles in the reconstituted ovary. When theca cells were labeled with CYP17a1, we found some overlapped staining with green fluorescent protein(GFP)-labeled BM-MSCs. The results suggest that BM-MSCs may participate in directing the differentiation of theca layer in the reconstituted ovary.ConclusionsThe presence of BM-MSCs in the artificial ovary was found to promote the survival of ovarian cells, as well as facilitate follicle formation and development. Since the cells didn’t proliferate in the reconstituted ovary, this discovery suggests a potential new and safe method for the application of MSCs in clinical fertility preservation by enhancing the success rate of cryo-thawed ovarian tissues after transplantation.

Inhibition of checkpoint kinase prevents human oocyte apoptosis induced by chemotherapy and allows enhanced tumour chemotherapeutic efficacy.

Could inhibition of the checkpoint kinase (CHEK) pathway protect human oocytes and even enhance the anti-tumour effects, during chemotherapy? CHEK inhibitors prevented apoptosis of human oocytes induced by chemotherapy and even enhanced the anti-tumour effects. CHEK inhibitors showed ovarian protective effects in mice during chemotherapy, while their role in human oocytes is unclear. This experimental study evaluated the ovarian reserve of young patients (120 patients) with cancer, exposed or not exposed to taxane and platinum (TP)-combined chemotherapy. Single RNA-sequencing analysis of human primordial oocytes from 10 patients was performed to explore the mechanism of oocyte apoptosis induced by TP chemotherapy. The damaging effects of paclitaxel (PTX) and cisplatin on human oocytes were also evaluated by culturing human ovaries in vitro. A new mouse model that combines human ovarian xenotransplantation and patient-derived tumour xenografts was developed to explore adjuvant therapies for ovarian protection. The mice were randomly allocated to four groups (10 mice for each group): control, cisplatin, cisplatin + CK1 (CHEK1 inhibitor, SCH 900776), and cisplatin + CK2 (CHEK2 inhibitor, BML277). In the prospective cohort study, human ovarian follicles were counted and serum AMH levels were evaluated. RNA-sequencing analysis was conducted, and staining for follicular damage (phosphorylated H2AX histone; γH2AX), terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) assays and assessments of apoptotic biomarkers (western blot and immunofluorescence) were conducted in human ovaries. After the treatments, histological analysis was performed on human ovarian samples to investigate follicular populations, and oocyte damage was measured by γH2AX staining, BAX staining, and TUNEL assays. At the same time, the tumours were evaluated for volume, weight, and apoptosis levels. Patients who received TP chemotherapy showed decreased ovarian reserves. Single RNA-sequencing analysis of human primordial oocytes indicated that TP chemotherapy induced apoptosis of human primordial oocytes by causing CHEK-mediated TAp63α phosphorylation. In vitro culture of human ovaries showed greater damaging effects on oocytes after cisplatin treatment compared with that after PTX treatment. Using the new animal model, CHEK1/2 inhibitors prevented the apoptosis of human oocytes induced by cisplatin and even enhanced its anti-tumour effects. This protective effect appeared to be mediated by inhibiting DNA damage via the CHEK-TAp63α pathway and by generation of anti-apoptotic signals in the oocytes. N/A. This was a preclinical study performed with human ovarian samples, and clinical research is required for validation. These findings highlight the therapeutic potential of CHEK1/2 inhibitors as a complementary strategy for preserving fertility in female cancer patients. This work was financially supported by the National Natural Science Foundation of China (nos. 82001514 and 81902669) and the Fundamental Research Funds for the Central Universities (2021yjsCXCY087). The authors declare no conflict of interest.

Thyroid hormone triiodothyronine does not protect ovarian reserve from DNA damage induced by X-ray and cisplatin.

Cancer therapy can induce premature ovarian insufficiency, necessitating methods for preserving fertility in female cancer patients. However, the only accepted clinical practice for doing so is cryopreservation of embryos, unfertilized ova, and ovarian tissue, despite potential options such as in vitro maturation of follicles. Therefore, considerable interest has arisen in fertoprotective agents, with research on rat ovarian granulosa cells suggesting that triiodothyronine (T3) regulates an anti-apoptosis mechanism that protects the ovarian reserve from paclitaxel-induced DNA damage. In this study, we used postnatal day 5 mouse ovary to confirm the existence of T3 thyroid hormone receptor (THR), as well as to investigate the potential protective effects of T3 against cisplatin- and X-ray-induced apoptosis. We also tested the potential anti-apoptotic effect of T3 in the breast cancer cell line MDA-MB-231. We treated cultured mouse ovaries with varying concentration of T3 and 4μM cisplatin and 0.2Gy X-ray. Real-time PCR, histological analysis, immunoblot analysis, and immunofluorescence were performed to assess the potential anti-apoptotic effects of T3. We confirmed that THR alpha and beta are expressed in the mouse ovary. T3 (0.1, 1, 10, 100nM, and 1µM) does not protect ovarian reserve from cisplatin- or X-ray-induced apoptosis or DNA damage. Similarly, it does not protect mouse granulosa cells and MDA-MB-231 cells from cisplatin- or X-ray-induced apoptosis. Our findings suggest that T3 is ineffective as a fertoprotective agent, and its candidacy as a potential agent to preserve fertility should be reconsidered.

P-483 Improving in vitro culture of isolated human ovarian primordial-primary follicles by adding adipose derived stem cells

Abstract Study question Could adipose derived stem cells (ADSc) secrete survival- and growth-promoting factors able to improve in vitro culture (IVC) of human isolated follicles? Summary answer Co-encapsulation of isolated human primordial-primary follicles with ADSc enhances follicular survival, growth and maturation after 7 days of IVC. What is known already The ability to obtain in vitro mature oocyte from earliest follicular stage may help to preserve fertility in female cancer patients. Unfortunately, human primordial follicles show low rates of growth in vitro. The only study that obtained mature human oocytes from primordial follicle used an IVC multi-step method with a success rate lower than 1% and abnormal polar body. Recently, several studies have shown that, in mice, ADSc promote survival, antrum formation and oocyte maturation in vitro by secreting factors important for follicular development. To our knowledge, no IVC studies on human ovarian follicles have been attempted using ADSc. Study design, size, duration Fragments of frozen-thawed human ovarian tissue (n = 5) have been mechanically and enzymatically digested to isolate primordial-primary follicles. Eight-to-ten isolated unilaminar follicles have been embedded in alginate-matrigel, half with 20.000 ADSc and half alone. Alginate beads were then cultured in vitro for 7 days. At day (D) 7, we compared isolated follicles co-cultured with ADSc to isolated follicles cultured alone. Participants/materials, setting, methods Every two days, IVC medium has been collected to perform hormonal (estradiol, progesteron, anti-Müllerian hormone, androgen, dehydroepiandrosterone) and growth factors measurement (VEGF, bFGF, IGF, HGF) by enzyme-linked immunosorbent assay (ELISA) to study follicle and ADSc interactions. Follicle diameter have been measured at D0 and at D7 of IVC. At D0 and at the end of the IVC (D7), follicle viability (live dead assay) and ultrastructure (transmission electron microscopy [TEM]) have been investigated. Main results and the role of chance A total of 373 unilaminar follicles were obtained after enzymatic isolation. At the end of IVC the percentage of viable follicles (with <10% of dead granulosa cells) was found to be significantly higher in follicles co-cultured for 7 days with ADSc (63.3%) compared to follicles in vitro cultured alone (45.5%) (p < 0.01). A significant increase in follicle size was observed in both groups after culture (p < 0.0001). This increase was statistically higher in follicles co-cultured with ADSc than follicles cultured alone (p < 0.005), reflecting better follicle growth in the follicle-ADSc group. Preliminary results by TEM show ADSc did not affect the ultrastructure of isolated follicles after 7 days of IVC. Limitations, reasons for caution As demonstrated by our results, ASCs appears to improve IVC conditions and boost follicle activation and survival preserving the follicle ultrastructure. Our study, however, did not investigate the underlying mechanisms behind this improved follicle growth and survival after IVC. Wider implications of the findings Co-encapsulation of human unilaminar follicles with ADSCs promotes early-stage ovarian follicle development and survival, possibly because of bidirectional interactions between follicles and ADSCs. This represents a promising first step for fertility preservation in female cancer patients who cannot benefit from cryopreserved ovarian tissue transplantation. Trial registration number not applicable

The cyto-protective effects of LH on ovarian reserve and female fertility during exposure to gonadotoxic alkylating agents in an adult mouse model.

STUDY QUESTIONDoes LH protect mouse oocytes and female fertility from alkylating chemotherapy?SUMMARY ANSWERLH treatment before and during chemotherapy prevents detrimental effects on follicles and reproductive lifespan.WHAT IS KNOWN ALREADYChemotherapies can damage the ovary, resulting in premature ovarian failure and reduced fertility in cancer survivors. LH was recently suggested to protect prepubertal mouse follicles from chemotoxic effects of cisplatin treatment.STUDY DESIGN, SIZE, DURATIONThis experimental study investigated LH effects on primordial follicles exposed to chemotherapy. Seven-week-old CD-1 female mice were randomly allocated to four experimental groups: Control (n = 13), chemotherapy (ChT, n = 15), ChT+LH-1x (n = 15), and ChT+LH-5x (n = 8). To induce primary ovarian insufficiency (POI), animals in the ChT and ChT+LH groups were intraperitoneally injected with 120 mg/kg of cyclophosphamide and 12 mg/kg of busulfan, while control mice received vehicle. For LH treatment, the ChT+LH-1x and ChT+LH-5x animals received a 1 or 5 IU LH dose, respectively, before chemotherapy, then a second LH injection administered with chemotherapy 24 h later. Then, two animals/group were euthanized at 12 and 24 h to investigate the early ovarian response to LH, while remaining mice were housed for 30 days to evaluate short- and long-term reproductive outcomes. The effects of LH and chemotherapy on growing-stage follicles were analyzed in a parallel experiment. Seven-week-old NOD-SCID female mice were allocated to control (n = 5), ChT (n = 5), and ChT+LH-1x (n = 6) groups. Animals were treated as described above, but maintained for 7 days before reproductive assessment.PARTICIPANTS/MATERIALS, SETTING, METHODSIn the first experiment, follicular damage (phosphorylated H2AX histone (γH2AX) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay), apoptotic biomarkers (western blot), and DNA repair pathways (western blot and RT-qPCR) were assessed in ovaries collected at 12 and 24 h to determine early ovarian responses to LH. Thirty days after treatments, remaining mice were stimulated (10 IU of pregnant mare serum gonadotropin (PMSG) and 10 IU of hCG) and mated to collect ovaries, oocytes, and embryos. Histological analysis was performed on ovarian samples to investigate follicular populations and stromal status, and meiotic spindle and chromosome alignment was measured in oocytes by confocal microscopy. Long-term effects were monitored by assessing pregnancy rate and litter size during six consecutive breeding attempts. In the second experiment, mice were stimulated and mated 7 days after treatments and ovaries, oocytes, and embryos were collected. Follicular numbers, follicular protection (DNA damage and apoptosis by H2AX staining and TUNEL assay, respectively), and ovarian stroma were assessed. Oocyte quality was determined by confocal analysis.MAIN RESULTS AND THE ROLE OF CHANCELH treatment was sufficient to preserve ovarian reserve and follicular development, avoid atresia, and restore ovulation and meiotic spindle configuration in mature oocytes exposed at the primordial stage. LH improved the cumulative pregnancy rate and litter size in six consecutive breeding rounds, confirming the potential of LH treatment to preserve fertility. This protective effect appeared to be mediated by an enhanced early DNA repair response, via homologous recombination, and generation of anti-apoptotic signals in the ovary a few hours after injury with chemotherapy. This response ameliorated the chemotherapy-induced increase in DNA-damaged oocytes and apoptotic granulosa cells. LH treatment also protected growing follicles from chemotherapy. LH reversed the chemotherapy-induced depletion of primordial and primary follicular subpopulations, reduced oocyte DNA damage and granulosa cell apoptosis, restored mature oocyte cohort size, and improved meiotic spindle properties.LARGE SCALE DATAN/A.LIMITATIONS, REASONS FOR CAUTIONThis was a preliminary study performed with mouse ovarian samples. Therefore, preclinical research with human samples is required for validation.WIDER IMPLICATIONS OF THE FINDINGSThe current study tested if LH could protect the adult mouse ovarian reserve and reproductive lifespan from alkylating chemotherapy. These findings highlight the therapeutic potential of LH as a complementary non-surgical strategy for preserving fertility in female cancer patients.STUDY FUNDING/COMPETING INTEREST(S)This study was supported by grants from the Regional Valencian Ministry of Education (PROMETEO/2018/137), the Spanish Ministry of Science and Innovation (CP19/00141), and the Spanish Ministry of Education, Culture and Sports (FPU16/05264). The authors declare no conflict of interest.

Cryopreservation Options to Preserve Fertility in Female Cancer Patients: Available Clinical Practice and Investigational Strategies from the Oncology Guidelines Point of View

In recent years, advances in cancer treatment have improved the survival rate of cancer patients significantly. However, destructive damage to ovaries due to the therapies or cancer itself can cause different degrees of infertility in women of reproductive age that can affect their quality of life seriously. In this study, fertility cryopreservation options for female cancer patients in oncology guidelines were reviewed. Cryopreservation methods have a long history in reproductive biology and oncology. However, embryo and oocyte cryopreservation were the eligible restoration strategies in clinical oncology practice. Ovarian tissue cryopreservation (OTC) is the latest option recommended for fertility preservation in pre-pubertal and adult patients who cannot delay their treatment or in whom taking IVF hormones may have adverse effects on their cancer. Reports show that frozen-thawed ovarian tissue transplantation has led to more than 130 live births so far in patients, most of whom were cancer patients. Although OTC is indeed generally recognized as an investigational method, it is recommended in some important guidelines, such as ASCO 2018. Therefore, based on many clinical pieces of evidence , it is predicted that the investigational label will soon be removed, and OTC might be considered as one of the main fertility preservation options for female cancer patients in clinical oncology practice.

Melatonin provides protection against cisplatin-induced ovarian damage and loss of fertility in mice

Research questionCan melatonin provide non-invasive ovarian protection against damage caused by cis-diamminedichloroplatinum (cisplatin) and preserve fertility in female cancer patients? And if so, what is the possible mechanism? DesignAthymic BALB/c nude tumour-bearing female mice were used to demonstrate whether melatonin affects the antineoplastic effect when co-administrated with cisplatin. Sexually mature and newborn C57BL/6 female mice were used to evaluate the potential effects of melatonin on the ovarian follicle pool, pregnancy rate and litter number in cisplatin-treated mice. The ovaries underwent immunohistochemical, TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) and gene array analysis to explore the underlying mechanism. In addition, granulosa cells were isolated to investigate the potential protective mechanism of melatonin. ResultsMelatonin not only enhanced the anti-cancer effect of cisplatin in tumour-bearing nude mice, but also reduced ovarian toxicity and preserved long-term fertility in cisplatin-treated C57BL/6 female mice. When co-administrated, melatonin was able to reduce the DNA damage and toxic effects on lipid peroxidation in the ovaries caused by cisplatin. Specifically, melatonin was able to largely restore lipid peroxidation in granulosa cells and thus prevent ovarian follicles from being depleted. ConclusionsMelatonin has the potential to be used as a chemotherapeutic adjuvant to simultaneously improve the outcome of anti-cancer treatment and preserve ovarian function during cisplatin chemotherapy. Notably, its properties of DNA protection and antioxidant effects on follicles may benefit female cancer survivors and prevent premature ovarian failure as well as fertility loss caused by chemotherapy.

Antioxidative effect of melatonin on cryopreserved ovarian tissue in mice

Cryopreservation of ovarian tissues (OTs) has become the most effective way to preserve the fertility of female cancer patients. However, cryopreservation of OTs is still relatively at an experimental stage. The aim of study is to examine the effect of melatonin (MTL) on cryopreserved-thawed OTs. Fragments of OTs were cryopreserved in medium containing different concentrations (0 mM, 0.001 mM, 0.01 mM, 0.1 mM and 1 mM) of MLT. The endogenous enzymes (GSH-PX, GSH, SOD, CAT and T-AOC), MDA and ROS levels were all evaluated after cryopreservation. Our results showed that the 0.1 mM of MLT significantly improved the survival and diameter of follicles (P < 0.001). Meanwhile, the antioxidant enzymes activities (including GSH-PX, GSH, SOD, CAT and T-AOC) were enhanced and MDA content were significantly decreased in 0.1 mM of MLT group compared to other groups (P < 0.001). Additionally, compared to the control group, MTL of 0.1 mM resulted in a significantly lower ROS level. In conclusion, MLT protects the quality of cryopreserved OTs by decreasing oxidative stress level and the optimal concentration is 0.1 mM.

Impacto dos agentes antineoplásicos sobre os folículos ovarianos e importância das biotécnicas reprodutivas na preservação da fertilidade humana

The preservation of fertility in young women of reproductive age has become a major challenge in medicine, since most cancer treatments can cause premature ovarian failure due to ovarian toxicity of chemotherapeutic agents. Based on this sensitivity it is necessary to revert or minimize damage to reproductive cells resulting from chemotherapy. It have become more frequent studies and researches for alternatives to preserve female fertility, and treatments that are more selective for cancer cells. Currently there are several options for preservation of fertility in women undergoing chemotherapy treatments.This article aims to present an overview of the damage caused by chemotherapy ovarian function and possible options for preserving fertility in female cancer patients and prospects in oncofertilidade. A search on databases were searched for relevant articles: the National Library of Medicine (PubMed) and the Virtual Health Library. Articles published between 1967 and 2015 on chemotherapeutic agents and their damage on female fertility was held.

Preserving fertility in female cancer patients: A snapshot of the options

With the odds of surviving cancer improving, many young women facing chemotherapy or radiotherapy may first wish to take steps to ensure that they will still be able to bear children afterward. The options depend on the type of disease, the treatment required, the age of the patient, whether she has a long-term partner, and whether cancer treatment can be delayed. This paper is an overview of current and experimental strategies for preserving fertility in female cancer patients.

Preservation of Fertility in Female Cancer Patients Desiring Future Child Bearing; What is Available and What can be Offered.

The aim of this review is to present an overview of available methods for preservation of ovarian function and fertility in female cancer patients who desire to maintain their child-bearing capacity for future pregnancies. A Medline search was conducted. Published articles from American and European studies from 1976 to present were reviewed. The effect of cancer treatment on the ovary, as well as different methods of fertility preservation and their reproductive outcomes are presented. Pregnancy rates vary according to the type of primary malignancy, stage of disease, method of fertility preservation (for example, hormonal therapy, cryopreservation, fertility-sparing surgery), and other confounding factors such as the patient’s age, reproductive capacity, status of partnership, and genetic disposition. The highest rates of successful pregnancy were observed with embryo cryopreservation. Today, higher cure rates and longer survival are a result of earlier cancer diagnosis and treatment. In conjunction with the advances in assisted reproduction, the preservation of ovarian function and fertility is a major part of multidisciplinary care that should be offered to any young female patient with cancer. Fertility preservation in young cancer patients raises a number of ethical issues particularly regarding standard versus experimental therapies, and long-term financial cost.

Retraction Note to: Successful cryopreservation of human ovarian cortex tissues using supercooling

The development of new method to cryopreserve human ovarian cortex tissues without damage is needed for the improvement of quality of life (QOL) of female cancer patients. Here we show novel cryopreservation method of human ovarian cortex tissues by using supercooling (S.C.) procedure. Our method will be helpful in order to preserve fertility of female cancer patients.

Successful cryopreservation of human ovarian cortex tissues using supercooling

The development of new method to cryopreserve human ovarian cortex tissues without damage is needed for the improvement of quality of life (QOL) of female cancer patients. Here we show novel cryopreservation method of human ovarian cortex tissues by using supercooling (S.C.) procedure. Our method will be helpful in order to preserve fertility of female cancer patients.

World of Reproductive Biology

A study in mice points to a new approach to preserving fertility in female cancer patients. Chemotherapy can wreak havoc on DNA. The drugs can damage the DNA of the oocyte pool, resulting in the induction of a cell-death program and culling of primordial oocytes, which provide a lifetime’s supply of eggs. A consequence of cancer therapy in women is often reduction of the ovarian reserve, resulting in impaired fertility and early menopause. Approaches to fertility preservation currently rely mainly on freezing oocytes or ovarian tissue prior to chemotherapy. But reproductive biologists have been seeking ways to protect oocytes from the effects of the chemotherapeutic agents by pharmacological means. The new study edges researchers closer to this goal. In this investigation, Jeffrey Kerr, Karla Hutt, Ewa Michalak, and colleagues took a close look at the triggers for oocyte death in response to DNAdamaging agents. Consistent with previous results, they found that DNA damage induces the activity of the transcriptional regulator TAp63, a homolog of Trp53—also known as p53, the famed ‘‘guardian of the genome.’’ The researchers also showed that TAp63 induced the expression of two factors that promote apoptosis, Puma and Noxa (both are members of the BH3-only pro-apoptotic subgroup of the wider BCL-2 family). Crucially, they showed that the primordial oocytes in mice lacking Puma or both Puma and Noxa were protected from apoptosis induced by DNA damage. These females could also produce apparently healthy offspring with no overt deformities and normal fertility. The mice lacking these genes also seemed to have active DNA repair pathways: A protein involved in double-stranded break repair (cH2AX) accumulated on damaged oocyte DNA in the oocytes of these mice, but was largely absent after five days, suggesting that the repair protein had done its job. It’s unclear whether the offspring suffer from damage to their DNA that might yield more subtle effects, and the researchers acknowledge the need for longer-term follow-up studies. If the same mechanism operates in humans, drugs that block PUMA and NOXA may be useful for preventing infertility in female cancer patients and could possibly even fend off menopause in healthy women. Kerr JB, Hutt KJ, Michalak EM, Cook M, Vandenberg CJ, Liew SH, Bouillet P, Mills A, Scott CL, Findlay JK, Strasser A. DNA damageinduced primordial follicle oocyte apoptosis and loss of fertility require TAp63-mediated induction of Puma and Noxa. Mol Cell 2012; 48(3):343–352.

A new in vitro model for pre-transplantation diagnosis of frozen/thawed human ovarian tissue

Introduction: Ovarian tissue freezing and autotransplantation is a promising option for preserving fertility of female cancer patients. There are several in vivo, but no established in vitro models to evaluate frozen/thawed human ovarian tissue prior to autotransplantation. Objective: To establish a new in vitro model for endocrine assessment of frozen/thawed human ovarian tissue prior to autotransplantation. Materials & Methods: Ovarian tissues were obtained by laparoscopy from 5 patients 19 to 34 years old recently diagnosed with cancer. 200mg cortical tissue from each patient was used and further divided into two groups: 100mg tissue as fresh (control group) and 100mg tissue as frozen/thawed (study group) after using slow freezing/rapid thawing protocol. Either fresh or frozen/thawed tissue from each patient was separately cultured in blank G-MOPS medium (Vitrolife, Sweden) for 6 days, at 37°C in an incubator without CO2. Estradiol (E2), Progesterone (P) and Anti Müllerian Hormone (AMH) were measured every 2 days in all supernatants to evaluate the endocrine function. After culture, all cortical tissues were examined histologically for viability and follicle development assessment. Mean values±SD were calculated. Results: Frozen/thawed tissues (study groups) showed slower recovery of endocrine function. On days 2, 4 and 6 of culture, E2 was 0.5±1.3, 4.4±9.8 & 14±13.3 pg/ml, P was 0.2±0, 0.2±0 & 0.3±0.1ng/ml, and AMH was below the detection limit. Viability after culture was 72±8.36%. Fresh tissues (control groups) showed faster recovery of endocrine function. On days 2, 4 and 6 of culture, E2 was 37.5±33.8, 62±44.7 & 104.2±58.9 pg/ml, P was 0.4±0.3, 0.6±0.4 & 1±0.5ng/ml, and AMH was under the detection limit. Viability after culture was 94±5.47%. Conclusion: For each patient, reasonable endocrine and histological data can be obtained by culturing only 100mg of fresh or frozen/thawed cortical tissue in G-MOPS medium for just 6 days. This new in vitro model can be used for each patient prior to autotransplantation to assess the endocrine function of the frozen/thawed ovarian tissue. These data will be complimented by follicle density assessment to confirm the endocrine results. In addition, oocyte isolation and in vitro maturation will be performed to assess the reproductive function.

In vivo delivery of FTY720 prevents radiation-induced ovarian failure and infertility in adult female nonhuman primates

To determine whether sphingosine-1-phosphate (S1P), or the S1P mimetic FTY720 shields ovaries of adult female rhesus monkeys from damage caused by 15 Gy of targeted radiotherapy, allowing for the retention of long-term fertility, and to evaluate whether S1P protects human ovarian tissue (xenografted into mice) from radiation-induced damage. Research animal study. Research laboratory and teaching hospital. Adult female rhesus macaques (8-14 years of age; n = 21) and two women (24 and 27 years of age) undergoing gynecologic surgery for benign reasons, after informed consent and approval. None. Ovarian histologic analysis, ovarian reserve measurements, and fertility in mating trials. Rapid ovarian failure was induced in female macaques by ovarian application of 15 Gy of radiation. Females given S1P or FTY720 by direct intraovarian cannulation for 1 week before ovarian irradiation rapidly resumed menstrual cycles because of maintenance of follicles, with greater beneficial effects achieved using FTY720. Monkeys given the S1P mimetic before ovarian irradiation also became pregnant in mating trials. Offspring conceived and delivered by radioprotected females developed normally and showed no evidence of genomic instability, as measured by micronucleus frequency in reticulocytes. Adult human ovarian cortical tissue xenografted into mice also exhibited a reduction in radiation-induced primordial oocyte depletion when preexposed to S1P. S1P and its analogs hold clinical promise as therapeutic agents to preserve ovarian function and fertility in female cancer patients exposed to cytotoxic treatments.

Medical approaches to preservation of fertility in female cancer patients

Introduction: The loss or impairment of ovarian function is an irreversible side effect that can occur in young cancer patients undergoing anticancer treatments. Its incidence varies according to the type of chemotherapy and the patient's age.Areas covered: The review includes studies or data available in literature from 1987 to 2010, examining current strategies to protect ovarian function and/or fertility in patients undergoing chemotherapy, which include oocyte, embryo or ovarian tissue cryopreservation, and temporary ovarian suppression during chemotherapy obtained by the administration of gonadotropin-releasing hormone analogues (GnRHa). The reader will gain an understanding of the incidence of premature ovarian function loss associated with chemotherapy; the advantages and disadvantages of the different strategies in protecting ovarian function; and the magnitude of the effect of GnRHa strategy in preserving ovarian function during chemotherapy.Expert opinion: The administration of GnRHa before and during chemotherapy is associated with an absolute reduction in the incidence of early menopause of nearly 20%. Such a strategy may be offered to young cancer patients who are candidates for chemotherapy. The capability of such an approach in inducing long-term preservation of ovarian function including fertility is still unknown.

In Vitro Maturation of Oocytes as a Strategy for Fertility Preservation

In vitro fertilization and embryo cryopreservation are regarded as the only established method for the preservation of fertility in female cancer patients. However, a possible delay in the treatment of cancer and exposure to supraphysiologic estrogen levels caused by ovarian stimulation raise concerns for patients and physicians. In vitro maturation avoids treatment delay or exposure to increased estradiol levels associated with in vitro fertilization. In vitro maturation combined with embryo or oocyte vitrification provides options that have been unavailable earlier, such as immature oocyte collection in the luteal phase, for some patients and improves the services provided by a fertility preservation program.

Preservation of Fertility in Female Cancer Patients

In this volume of Deutsches Arzteblatt, a group from the University Department of Gynecology at Erlangen report Germany’s first case of the reimplantation of cryopreserved ovarian tissue. This marks a step forward in efforts to preserve fertility for patients undergoing cancer treatments. Strategies to preserve fertility in cancer patients are much needed. The numbers of long-term cancer survivors especially in childhood, adolescence, and early adulthood are rising constantly. The five-year survival rate for Hodgkin’s lymphoma in childhood, for example, is over 90%, and almost 80% for acute lymphoblastic leukemia (1). In the year 2010, one in every 715 adults will have a history of childhood cancer (2). These good long-term prospects open the possibility of life choices, of which the choice to have a family is one. This is however compromised by the fact that the cytotoxic damage to the gonads resulting in infertility is a predictable consequence of oncological treatment, whether chemotherapy or radiotherapy.

Fertilitätsprotektion bei onkologischen Patientinnen: Kryobank Bonn - Beratung, Anwendung und die Bedeutung für die betroffene Frau

Fragestellung: Der Fertilitätsprotektion bei Krebspatientinnen wird nicht immer ausreichend Aufmerksamkeit zuteil. Ursachen dafür können ärztliche Unsicherheit sein, Patientinnen nach einer Krebsdiagnose mit diesem Thema nicht zusätzlich belasten zu wollen, außerdem die Einschätzung, wenig erfolgreiche Therapien zur Verfügung zu haben. Bislang ist wenig über die psychischen Aspekte von fertilitätserhaltenden Maßnahmen bekannt. Methode: In einer offenen Pilotstudie wurden bislang 30 Krebspatientinnen, die die Kryokonservierung von Ovargewebe in der Kryobank Bonn vorgenommen haben, hinsichtlich ihrer Motive, Erfahrungen und Verbesserungsvorschlägen bezüglich der Kryokonservierung von Ovargewebe schriftlich befragt. Ergebnisse: Teilnehmende Frauen waren hauptsächlich am Hodgkin-Lymphom, Mamma-Ca oder an Darmkrebs erkrankt. Auffallend war der hohe Bildungsstand (50 % Hochschulstudium). Die meisten Frauen waren zufrieden mit der Aufklärung zur Kryokonservierung, einige merkten kritisch an, dass der Informationsstand vorbehandelnder Gynäkologen bzw. Onkologen hinsichtlich Fertilitätsprotektion defizitär sei. Motive für die Inanspruchnahme der Kryokonservierung von Ovargewebe war für alle Frauen ein zumindest theoretischer Kinderwunsch. Frauen waren sich bewusst, dass die Kryokonservierung keine Garantie für eine erfolgreiche Schwangerschaft gibt, jedoch war für sie das Gefühl zentral, „jede Möglichkeit ausgeschöpft zu haben“. Über die Hälfte der Patientinnen beschrieben einen positiven Einfluss ihrer Entscheidung zur Kryokonservierung auf das Krankheitserleben. Schlussfolgerung: Gynäkologen und Onkologen sollten Krebspatientinnen im reproduktiven Alter über die Möglichkeiten des Fertilitätsverlustes und ‐erhaltes aufklären und an spezialisierte Zentren verweisen. Vor dem Hintergrund der positiven Auswirkung auf die Krankheitsbewältigung ist der Einsatz der Kryokonservierung von Ovargewebe positiv zu bewerten - auch wenn das Verfahren hinsichtlich der Schwangerschaftsraten noch als experimentell bewertet wird.