Fertility Preservation Program Research Articles

P-345 Too cool to be late: the discrepancy between the required and obtained number of oocytes in a social fertility preservation program

Abstract Study question What are the outcomes of thawed oocytes from social fertility preservation (SFP) programs by age? Is the number of cryopreserved oocytes sufficient to achieve pregnancy? Summary answer The outcomes of thawed oocytes from SFP programs diminish with age, and the number of cryopreserved oocytes is below the desirable quantity for pregnancy achievement. What is known already Social fertility preservation empowers women to safeguard their fertility by anticipating age-related decline and diminished efficacy of fertility treatments in older ages. The effectiveness of oocyte vitrification programs, and consequently fertility preservation, depends on the patient’s age. Previous reports on fertility preservation indicate that, typically, a larger number of oocytes are retrieved in patients aged ≤35 when compared to older patients. Although these reports document that oocyte vitrification preserves developmental potential akin to fresh oocytes, uncertainties persist regarding patient awareness of the adequate number of cryopreserved oocytes needed for pregnancy. Study design, size, duration This historical cohort study, conducted at a university-affiliated IVF center from January 2012 to December 2022, analyzed data from 1,332 vitrified oocyte ICSI cycles involving 1,000 SFP program participants. Fertility preservation patients and oocyte thawing cycles (n = 132) were categorized by age as follows: <30 years (n = 55 and 24), 30-35 years (n = 250 and 33), 36-40 years (n = 480 and n = 42), and >40 years (n = 215 and 33) for preservation and thawing, respectively. Participants/materials, setting, methods Laboratory and clinical outcomes were compared among the groups. In a subsequent analysis, data from PGT cycles conducted in our laboratory (n = 480) from January to December 2022 were utilized to determine blastulation and euploidy rates within vitrified oocytes. This data, combined with literature findings, was employed to estimate the average number of oocytes required for a patient to reach a 95% cumulative probability of pregnancy. These estimates were then compared with our database findings. Main results and the role of chance There were 1,332 cycles of SFP involving 1,000 patients. Among them, 121 (12.2%) returned for oocyte thawing and underwent 132 ICSI cycles. A significant difference in the survival rate was observed among thawed oocytes from patients under 30 compared to those over 40 years-old (<30: 90.7 ± 33, 30-35: 81.4 ± 40.4, 36-40: 83.4 ± 35.5, and >40: 72.3 ± 34, p = 0.047). Additionally, a significantly higher pregnancy rate was observed in the 30–35 age group when compared to those in the group aged >40 years-old (< 30: 50.0 ± 17.7, 30-35: 56.0 ± 85.0, 36-40: 46.3 ± 69.0, and >40: 21.4 ± 93.0, p = 0.048). However, there was no significant difference when comparing fertilization, blastulation, and implantation rates. In the general group, an appropriate number of oocytes to be cryopreserved to achieve a pregnancy chance of 95% would be 17. For patients aging < 30, 30-35, 36-40, and > 40, that would be 7.47, 10.7, 22.5, and 42.8, respectively. Overall, 92 out of 1,000 patients (9.2%) obtained an adequate number of oocytes. When the group was stratified by age, it was observed that 54.5%, 27.2%, 2.5%, and 0% of the patients obtained an adequate number of oocytes to achieve pregnancy. Limitations, reasons for caution The retrospective nature of the study and the small patient cohort included in the analysis, when divided by age, may pose limitations. However, the low number of patients returning to utilize their oocytes in SFP is a consistent limitation across all studies. Wider implications of the findings These findings underscore the importance of appropriate counseling and encouragement for patients undergoing SFP earlier, ensuring a larger number of oocytes are cryopreserved. This aligns with SFP’s primary objective: enhancing women’s reproductive autonomy by enabling the postponement of childbearing and preserving the biological relationship with future offspring. Trial registration number N/A

Fertility Preservation in the Era of Immuno-Oncology: Lights and Shadows.

In recent years, immuno-oncology has revolutionized the cancer treatment field by harnessing the immune system's power to counteract cancer cells. While this innovative approach holds great promise for improving cancer outcomes, it also raises important considerations related to fertility and reproductive toxicity. In fact, most young females receiving gonadotoxic anti-cancer treatments undergo iatrogenic ovarian exhaustion, resulting in a permanent illness that precludes the vocation of motherhood as a natural female sexual identity. Although commonly used, oocyte cryopreservation for future in vitro fertilization and even ovarian cortex transplantation are considered unsafe procedures in cancer patients due to their oncogenic risks; whereas, ovarian stem cells might support neo-oogenesis, providing a novel stemness model of regenerative medicine for future fertility preservation programs in oncology. Recent scientific evidence has postulated that immune checkpoint inhibitors (ICIs) might in some way reduce fertility by inducing either primary or secondary hypogonadism, whose incidence and mechanisms are not yet known. Therefore, considering the lack of data, it is currently not possible to define the most suitable FP procedure for young patients who are candidates for ICIs. In this report, we will investigate the few available data concerning the molecular regulation of ICI therapy and their resulting gonadal toxicity, to hypothesize the most suitable fertility preservation strategy for patients receiving these drugs.

Expansion of the Fertility Preservation Program to All Newly Diagnosed Prepubertal Patients with Cancer at a Pediatric Hospital.

Background: As the majority of pediatric patients with cancer survive their disease, generating a population of over 500,000 childhood cancer survivors in the United States, it is imperative to minimize the lifelong consequences of treatment, which include temporary or permanent infertility caused by certain cancer treatments. A fertility consultation at diagnosis can provide patients and families with the opportunity to be informed regarding the likelihood of gonadal dysfunction and to consider fertility preservation. Method: After our pediatric hospital started to offer tissue cryopreservation, we initiated this evidence-based interventional quality improvement project. Our primary aim was to ensure that all newly diagnosed prepubertal patients with cancer who met the criteria for fertility tissue preservation were correctly identified and offered an educational consultation and preservation. Results: Between July 15, 2022 and October 30, 2022, 54 patients' treatment plans were evaluated to determine treatment-related infertility risk using the Oncofertility Consortium Pediatric Initiative Network's Risk Assessment tool. Fifteen patients were at a high level of significantly increased risk and 13 were eligible for consultation. Seven (46%) patients and their families received a consultation. Initiation of treatment before referral was the primary reason for lack of consultation. Six of seven patients receiving consultation (86%) elected to undergo preservation. Preservation procedures did not cause a delay in starting treatment for those patients. Discussion: A fertility preservation program with established policies and processes can increase the likelihood that prepubertal patients at high risk for infertility are correctly identified, educated, and offered preservation.

“Being useful, I think it's the result of a sick society”: Critical reflections on reproductive politics and markets by women freezing their eggs in Spain

AbstractThis piece analyzes the way in which women that froze, are considering freezing or are freezing their eggs in Spain think critically about broader reproductive politics in Spain and about assisted reproduction. Drawing partially on previous studies around egg freezing, Thomas Lemke has suggested that cryopreservation practices represent a “politics of suspension” characterized by both reversibility and disposition, and concomitant with broader political inaction (Lemke in Sci Technol Hum Values 48(4):1–27, 2021). Drawing on feminist literature, and on how some of these women think about motherhood, it is relevant to emphasize this ‘suspension of politics’ that takes place along with a “politics of suspension,” meaning that certain matters (such as reproduction and its postponement) are only to be dealt with privately and individually, through marketized fertility preservation programs in this case. Some of the women interviewed describe these programs as useful tools within a problematic context: technologies that give time in a context that leaves them on their own to figure out motherhood (or its absence) in the midst of uncertainty and loneliness. This paper shows their critical views on these matters, while reflecting on how their experiences and desires become increasingly imbricated with the fertility industry in the making of their reproductive biographies (Perler and Schurr in Body Soc 27(3): 3–27, 2021).

A dozen years of ovarian tissue cryopreservation at a pediatric hospital: tracking program and patient metrics while adapting to increasing needs

To review the program and patient metrics for ovarian tissue cryopreservation (OTC) within a comprehensive pediatric fertility preservation program in its first 12 years of development. Retrospective review. A tertiary children's hospital in a large urban center between March 2011 and February2023. Pediatric patients who underwent OTC. Unilateral oophorectomy for OTC. Patient demographics and clinical course information were collected for analysis. A total of 184 patients underwent OTC in the first 12 years. One hundred fifteen patients were prepubertal at the time of OTC, and 69 were postpubertal. In total, 128 patients (69.6%) received part of their planned therapy before OTC. Starting in 2018, 104 participants (92.0%) donated tissue to research, 99 participants (87.6%) donated blood, and 102 (90.2%) donated media to research. There was a decrease in the median age of patients who underwent OTC from 16.4-6.6 years and an overall increase in the proportion of patients per year that were prepubertal. Forty-eight (26.0%) patients who underwent OTC were outside referrals and traveled from as far as Seattle, Washington. During the first 12 years of this program, oncofertility research increased, annual tissue cryopreservation cases increased, and the median age of those who underwent OTC decreased. The program was adapted to build a stand-alone gonadal tissue processing suite and specialized in prepubertal ovarian tissue processing. The program will continue to adapt to patient needs in the upcoming decades because restoration technologies advance through research supported by this and collaborating programs.

A 20-year overview of fertility preservation in boys: new insights gained through a comprehensive international survey.

Twenty years after the inception of the first fertility preservation programme for pre-pubertal boys, what are the current international practices with regard to cryopreservation of immature testicular tissue? Worldwide, testicular tissue has been cryopreserved from over 3000 boys under the age of 18 years for a variety of malignant and non-malignant indications; there is variability in practices related to eligibility, clinical assessment, storage, and funding. For male patients receiving gonadotoxic treatment prior to puberty, testicular tissue cryopreservation may provide a method of fertility preservation. While this technique remains experimental, an increasing number of centres worldwide are cryopreserving immature testicular tissue and are approaching clinical application of methods to use this stored tissue to restore fertility. As such, standards for quality assurance and clinical care in preserving immature testicular tissue should be established. A detailed survey was sent to 17 centres within the recently established ORCHID-NET consortium, which offer testicular tissue cryopreservation to patients under the age of 18 years. The study encompassed 60 questions and remained open from 1 July to 1 November 2022. Of the 17 invited centres, 16 completed the survey, with representation from Europe, Australia, and the USA. Collectively, these centres have cryopreserved testicular tissue from patients under the age of 18 years. Data are presented using descriptive analysis. Since the establishment of the first formal fertility preservation programme for pre-pubertal males in 2002, these 16 centres have cryopreserved tissue from 3118 patients under the age of 18 years, with both malignant (60.4%) and non-malignant (39.6%) diagnoses. All centres perform unilateral biopsies, while 6/16 sometimes perform bilateral biopsies. When cryopreserving tissue, 9/16 centres preserve fragments sized ≤5 mm3 with the remainder preserving fragments sized 6-20 mm3. Dimethylsulphoxide is commonly used as a cryoprotectant, with medium supplements varying across centres. There are variations in funding source, storage duration, and follow-up practice. Research, with consent, is conducted on stored tissue in 13/16 centres. While this is a multi-national study, it will not encompass every centre worldwide that is cryopreserving testicular tissue from males under 18 years of age. As such, it is likely that the actual number of patients is even higher than we report. Whilst the study is likely to reflect global practice overall, it will not provide a complete picture of practices in every centre. Given the research advances, it is reasonable to suggest that cryopreserved immature testicular tissue will in the future be used clinically to restore fertility. The growing number of patients undergoing this procedure necessitates collaboration between centres to better harmonize clinical and research protocols evaluating tissue function and clinical outcomes in these patients. K.D. is supported by a CRUK grant (C157/A25193). R.T.M. is supported by an UK Research and Innovation (UKRI) Future Leaders Fellowship (MR/S017151/1). The MRC Centre for Reproductive Health at the University of Edinburgh is supported by MRC (MR/N022556/1). C.L.M. is funded by Kika86 and ZonMW TAS 116003002. A.M.M.v.P. is supported by ZonMW TAS 116003002. E.G. was supported by the Research Program of the Research Foundation-Flanders (G.0109.18N), Kom op tegen Kanker, the Strategic Research Program (VUB_SRP89), and the Scientific Fund Willy Gepts. J.-B.S. is supported by the Swedish Childhood Cancer Foundation (TJ2020-0026). The work of NORDFERTIL is supported by the Swedish Childhood Cancer Foundation (PR2019-0123; PR2022-0115), the Swedish Research Council (2018-03094; 2021-02107), and the Birgitta and Carl-Axel Rydbeck's Research Grant for Paediatric Research (2020-00348; 2021-00073; 2022-00317; 2023-00353). C.E is supported by the Health Department of the Basque Government (Grants 2019111068 and 2022111067) and Inocente Inocente Foundation (FII22/001). M.P.R. is funded by a Medical Research Council Centre for Reproductive Health Grant No: MR/N022556/1. A.F. and N.R. received support from a French national research grant PHRC No. 2008/071/HP obtained by the French Institute of Cancer and the French Healthcare Organization. K.E.O. is funded by the University of Pittsburgh Medical Center and the US National Institutes of Health HD100197. V.B-L is supported by the French National Institute of Cancer (Grant Seq21-026). Y.J. is supported by the Royal Children's Hospital Foundation and a Medical Research Future Fund MRFAR000308. E.G., N.N., S.S., C.L.M., A.M.M.v.P., C.E., R.T.M., K.D., M.P.R. are members of COST Action CA20119 (ANDRONET) supported by COST (European Cooperation in Science and Technology). The Danish Child Cancer Foundation is also thanked for financial support (C.Y.A.). The authors declare no competing interests. N/A.

Generation of two isogenic sickle cell disease induced pluripotent stem cell lines from testicular fibroblasts

Curative bone marrow transplantation (BMT) therapies for sickle cell disease (SCD) can cause infertility. The Fertility Preservation Program (FPP) in Pittsburgh cryopreserves testicular tissues for SCD patients prior to BMT in anticipation that those tissues can be thawed in the future and matured to produce sperm. Here, we generated and validated two isogenic patient-derived induced pluripotent stem cell (iPSC) lines from testicular biopsy fibroblasts of a 12-year-old SCD patient.

Current status of fertility preservation in a Spanish tertiary public hospital: multidisciplinary approach and experience in over 1500 patients.

Currently, 15% of gynaecological and 9% of haematological malignancies are diagnosed before the age of 40. The increased survival rates of cancer patients who are candidates for gonadotoxic treatments, the delay in childbearing to older ages, and the optimization of in vitro fertilisation techniques have all contributed to an increased interest in fertility preservation (FP) treatments. This study reviews the experience of the Fertility Preservation Programme (FPP) of a tertiary public hospital with a multidisciplinary approach. This retrospective study included all the available (FP) treatments, performed in patients of childbearing age between 2006 and 2022. 1556 patients were referred to the FPP: 332 oocyte vitrification cycles, 115 ovarian cortex cryopreservation with 11 orthotopic autotransplantations, 175 gonadotropin-releasing hormone (GnRH) agonist treatments, 109 fertility-sparing treatments for gynaecological cancer, and 576 sperm cryopreservation were performed. Malignancy was the main indication for FP (the main indications being breast cancer in women and haematological malignancies in men), although non-oncological pathologies, such as endometriosis and autoimmune diseases, have increased in recent years. Currently, the most widely used FP technique is oocyte vitrification, the increase of which has been associated with a decrease in the use of cortex CP and GnRH agonists. The increase in FP treatment reflects the implementation of reproductive counselling in oncology programmes. A multidisciplinary approach in a tertiary public hospital allows individualised FP treatment for each patient. In recent years, there has been a change in trend with the introduction of new indications for FP and a change in techniques due to their optimisation.

FGF2, LIF, and IGF1 (FLI) supplementation during human in vitro maturation enhances markers of gamete competence.

Does a chemically defined maturation medium supplemented with FGF2, LIF, and IGF1 (FLI) improve in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) obtained from children, adolescents, and young adults undergoing ovarian tissue cryopreservation (OTC)? Although FLI supplementation did not increase the incidence of oocyte meiotic maturation during human IVM, it significantly improved quality outcomes, including increased cumulus cell expansion and mitogen-activated protein kinase (MAPK) expression as well as enhanced transzonal projection retraction. During OTC, COCs, and denuded oocytes from small antral follicles are released into the processing media. Recovery and IVM of these COCs is emerging as a complementary technique to maximize the fertility preservation potential of the tissue. However, the success of IVM is low, especially in the pediatric population. Supplementation of IVM medium with FLI quadruples the efficiency of pig production through improved oocyte maturation, but whether a similar benefit occurs in humans has not been investigated. This study enrolled 75 participants between January 2018 and December 2021 undergoing clinical fertility preservation through the Fertility & Hormone Preservation & Restoration Program at the Ann & Robert H. Lurie Children's Hospital of Chicago. Participants donated OTC media, accumulated during tissue processing, for research. Participants who underwent OTC and include a pediatric population that encompassed children, adolescents, and young adults ≤22 years old. All participant COCs and denuded oocytes were recovered from media following ovarian tissue processing. IVM was then performed in either a standard medium (oocyte maturation medium) or one supplemented with FLI (FGF2; 40 ng/ml, LIF; 20 ng/ml, and IGF1; 20 ng/ml). IVM outcomes included meiotic progression, cumulus cell expansion, transzonal projection retraction, and detection of MAPK protein expression. The median age of participants was 6.3 years, with 65% of them classified as prepubertal by Tanner staging. Approximately 60% of participants had been exposed to chemotherapy and/or radiation prior to OTC. On average 4.7 ± 1 COCs and/or denuded oocytes per participant were recovered from the OTC media. COCs (N = 41) and denuded oocytes (N = 29) were used for IVM (42 h) in a standard or FLI-supplemented maturation medium. The incidence of meiotic maturation was similar between cohorts (COCs: 25.0% vs 28.6% metaphase II arrested eggs in Control vs FLI; denuded oocytes: 0% vs 5.3% in Control vs FLI). However, cumulus cell expansion was 1.9-fold greater in COCs matured in FLI-containing medium relative to Controls and transzonal projection retraction was more pronounced (2.45 ± 0.50 vs 1.16 ± 0.78 projections in Control vs FLIat 16 h). Additionally, MAPK expression was significantly higher in cumulus cells obtained from COCs matured in FLI medium for 16-18 h (chemiluminescence corrected area 621,678 vs 2,019,575 a.u., P = 0.03). Our samples are from human participants who exhibited heterogeneity with respect to age, diagnosis, and previous treatment history. Future studies with larger sample sizes, including adult participants, are warranted to determine the mechanism by which FLI induces MAPK expression and activation. Moreover, studies that evaluate the developmental competence of eggs derived from FLI treatment, including assessment of embryos as outcome measures, will be required prior to clinical translation. FLI supplementation may have a conserved beneficial effect on IVM for children, adolescents, and young adults spanning the agricultural setting to clinical fertility preservation. This work was supported by Department of Obstetrics and Gynecology startup funds (F.E.D.), Department of Surgery Faculty Practice Plan Grant and the Fertility & Hormone Preservation & Restoration Program at the Ann & Robert H. Lurie Children's Hospital of Chicago (M.M.L. and E.E.R.). M.M.L. is a Gesualdo Foundation Research Scholar. Y.Y.'s research is supported by the internal research funds provided by Colorado Center of Reproductive Medicine. Y.Y., L.D.S., R.M.R., and R.S.P. have a patent pending for FLI. The remaining authors have no conflict of interest. N/A.

Disease-Free Survival Does Not Differ According to Fertility Preservation Technique for Young Women With Breast Cancer

ABSTRACT Approximately 5% of breast cancer cases occur in women younger than 40 years, and the 5-year overall survival for women between the ages of 15 and 44 years is 90%. As the childbearing age increases, so too does the proportion of women who wish to achieve pregnancy after breast cancer. The conception rate among these women is 40% lower than the general population, as a result of advanced age and the gonadal toxicity of chemotherapy. Fertility preservation (FP) techniques enable young patients to freeze gametes before treatment; however, the most recommended method of FP is oocyte or embryo vitrification after controlled ovarian hyperstimulation (COH), which has limited safety data in breast cancer patients. This retrospective cohort study aimed to evaluate disease-free survival (DFS) in a large cohort of women with breast cancer who underwent FP techniques with or without COH. Patients with breast cancer between 18 and 43 years old referred for FP to 2 university hospitals between July 2013 and July 2019 were included. Women not included in the FP program after oncofertility counseling, those with stage 4 disease at diagnosis or history of other invasive cancer, or those with gonadotoxic treatment have been excluded. Patients were split into 2 groups, with the STIM group receiving at least 1 COH protocol using a gonadotropin-releasing hormone antagonist with or without an aromatase inhibitor and the no STIM group not undergoing ovarian stimulation. Patient characteristics, obstetric variables, and oncology characteristics were collected from medical records. Disease-free survival and overall survival were estimated using Kaplan-Meier plots and compared between the 2 groups by log-rank test. A total of 740 women (76.7%) who attended oncofertility consultations for breast cancer were included. Of these, 328 (44.3%) underwent at least 1 ovarian stimulation cycle, and 412 (55.7%) had an FP technique without exogenous FSH administration. The STIM group was significantly older and had lower values in ovarian reserve tests, and more women in the no STIM group underwent a mastectomy and neoadjuvant chemotherapy. The proportion of patients with hormone receptor-positive tumors was equal in both groups. Letrozole was coadministered with exogenous gonadotropin in 39.3% of women in the STIM group (Let-COH). Overall, 16.8% of patients had a tumor still present on oocyte retrieval after COH, of which 55% received a Let-COH protocol. Follow-up data were available for 80.9% (559/740) of patients. Cancer recurrence was documented in 87 patients (27 in STIM, 60 in no STIM), and DFS was not statistically different between the groups (log-rank test P = 0.09). Neither difference in DFS nor overall survival was observed in the Let-COH group compared with those undergoing COH after breast surgery. Kaplan-Meier estimates of overall survival at 4 years were 97.6% in the STIM and 93.6% in the no STIM groups, and after adjusting for prognostic parameters, the risk of death remained significantly lower in the STIM group (hazards ratio, 0.55 [0.35–0.85]). The results of this large retrospective cohort study demonstrate that the use of COH for FP patients with breast cancer before or after breast tumor surgery is safe; however, a longer duration of follow-up will provide a more definite safety profile.

Impact of prepubertal bovine ovarian tissue pre-freeze holding duration on follicle quality

Ovarian tissue cryopreservation prior to gonadotoxic treatment is the only recommended option for fertility preservation in prepubertal girls. Due to the technical complexity of this technique, limited number of centres across the world are equipped to offer the facility. Hence, the retrieved ovarian tissue needs to be maintained at hypothermic temperature (4 °C) for long time during shipment. The time taken between tissue retrieval and cryopreservation could influence the functionality of cells during fertility restoration. This study explored the tissue integrity and follicle quality of ovarian cortical slices subjected to pre-freeze holding for various time durations in vitro. Prepubertal bovine ovarian tissue from < 12 months old animals were handled at hypothermic holding (4 °C) for 0, 24, 48 and 72 h. The tissues were assessed for follicle viability through confocal analysis of live-dead labelled samples, and follicle quality and tissue integrity through histology. Results have shown that follicle viability, and overall follicle quality were not significantly affected at the end of 72 h hypothermic holding. Though, the observation reassures extended hypothermic holding prior to freezing, findings need to be validated in human tissue prior to use in clinical fertility preservation programs.

P-429 Transcriptomic heterogeneity within cortical ovarian follicle pool in child and adult

Abstract Study question What are the transcriptomic differences between child and adult ovarian follicles in the human cortex? Summary answer Our results suggest that human ovarian cortical follicles show marked differences before and after puberty, with the greatest differences being present in the secondary stage. What is known already The ovaries and their main functional structures, follicles, are important for determining the reproductive health. Follicle growth and maturation are controlled by multiple mechanisms, including the hypothalamus pituitary ovarian axis and local paracrine signalling. Although dormant primordial follicles are activated to grow even in infancy, all growing follicles before the first menarche are bound to undergo atresia. Follicle growth activation in infancy and adulthood remains poorly understood. Additionally, child and adult follicles behave differently during in vitro culture suggesting differing biology, raising many questions e.g., about their suitability for fertility preservation. Study design, size, duration Ovarian cortical tissues from children were collected through fertility preservation program at the Helsinki Children’s hospital (Finland). Families of underage patients signed an informed written consent. Adult ovarian tissue was collected from gender reassignment patients at the Helsinki University hospital and Karolinska university hospital after informed written consent. Follicles at different maturation stages were isolated from ovarian cortex for single follicle transcriptomic analysis. Freshly-fixed cortical biopsies (3x3x1 mm3) were used for validation of transcriptomic analysis. Participants/materials, setting, methods Single viable follicles (n = 120, 60/age group) were mechanically and enzymatically isolated from frozen-thawed ovarian tissue from four adults (age &amp;gt;18 yrs) and five children (age 1-11 yrs) and processed for RNA-sequencing. Individual follicles were lysed, prepared for RNA libraries following Smart-Seq2 protocol and sequenced using NextSeq 500 platform. After quality assessment, 109 follicles were analysed (Adult n = 54, Child n = 55). DDX4, AMH, FOXL2 and DAZL transcripts and proteins localization were evaluated via immunodetection and RNA-FISH assay. Main results and the role of chance Principal component analysis (PCA) of all sequenced follicles demonstrated an expected timely progression of their stages based on maturation, and an overall similarity between child and adult. Interestingly, PCA displayed two distinct clusters of follicles in both adult and child samples. The first group contained only growing follicles and, after differentially expressed gene (DEG) analysis, demonstrated lower expression of traditional oocyte markers (group 1). The second group consisted of primordial and growing follicles and had lower expression of traditional granulosa marker (group 2). Group-related differential expression of main oocyte (DDX4, DAZL) and granulosa (FOXL2, AMH) marker genes were validated in tissue sections with RNA-FISH. Downstream analyses focused on group 2 follicles. When child and adult follicles were compared to each other, the highest number of DEGs were found in the secondary stage (primordial n = 32, primary n = 14, secondary n = 387). Pathway analysis of DEG displays enrichment of oestrogen-related related pathways in adult secondary follicles. Furthermore, we analysed DEGs between the follicular stages per age group, finding high number of DEGs between primordial and primary follicles (Adult n = 1663, Child n = 1914). The majority of those genes are unique to the age group (common=512). Limitations, reasons for caution Adult ovarian tissue is obtained from gender reassignment patients, who receive androgen treatments prior to surgery. Child ovarian tissue was collected from fertility preservation programme where some patients were exposed to chemotherapy prior to tissue collection. Wider implications of the findings The differences between child and adult follicles encourage further investigation of the suitability of child ovarian tissue in current fertility preservation protocols. Understanding transcriptomic changes during early folliculogenesis and reasons for follicle degeneration can help tailor protocols for in vitro growth of follicles. Trial registration number not applicable

P-421 Return rates and outcome of fertility preservation in women with cervical cancer.

Abstract Study question What are the return rates and reproductive outcome after fertility preservation (FP) in women with cervical cancer (CC)? Summary answer Return rates after FP in women with CC seem to be comparable with those after FP for other indications. What is known already Cervical cancer is the fourth most common cancer among women. Approximately 42% of the affected women are under the age of 45, and many of them wish to preserve fertility at the time of diagnosis. Fertility sparing treatment modalities and FP with cryopreservation of oocytes, embryos or/and ovarian tissue can be offered for this purpose to carefully selected patients. These treatments are offered at Swedish academic centers within the health insurance coverage available to all citizens. To date, data on return rates, reproductive outcome and survival in women with cervical cancer undergoing FP are scarce. Study design, size, duration Prospective, single center study aiming to report long-term outcomes in women with CC who have versus have not undergone FP at Karolinska University Hospital between January 1st 1999 and September 30th 2018. Participants/materials, setting, methods During the study period, 74 women with CC received FP counseling at Karolinska University Hospital, and 52 of them women proceeded to FP. Data on return rates, reproductive outcomes and overall survival were extracted using the clinical registries. Main results and the role of chance By January 15th, 2023, 22 of 52 women with FP returned for a new fertility counselling or treatment, mean time between FP and return was 4 years (1-7 years). Among women with cryopreserved ovarian tissue (n = 40), 17 have been in contact with the clinic with fertility wish, 3 of them had re-transplantation of the tissue, but none of them achieved oocyte retrieval yet. Additionally, one woman is planned for re-transplantation in February 2023, and one woman is awaiting the decision. In two cases the decision not to re-transplant ovarian tissue was based on absence of endometrial growth following attempts of hormonal substitutive therapy. Among women with cryopreserved oocytes (n = 5), 1 returned for thawing but no pregnancy has been achieved. Among women with cryopreserved embryos (n = 5), 3 returned during follow-up, 2 proceeded to thawing and transfer, 1 got pregnant and gave birth to a child. Of two women with combination of oocytes and ovarian tissue cryopreserved, one returned for fertility treatment, but no pregnancy has been achieved after thawing and re-transplantation. Eight women died during follow-up because of recurrence of their cancer, 4 of 22 in the group without FP and 4 of 52 in the group with FP. Limitations, reasons for caution This study provides much needed data on real-world outcome in women with FP indicated by diagnosis of CC, but it has the limitations related to its descriptive character. The use of gestational carriers is not permitted in Sweden and FP program may differ from those in other countries. Wider implications of the findings The study results provide much needed data on real-world reproductive and oncologic outcome following FP in women with CC. Trial registration number NTC04602962

P-409 Oocyte cryopreservation following ovarian stimulation for fertility preservation in transgender men: a French case series

Abstract Study question To evaluate the feasibility and results of a fertility preservation program for transgender men. Summary answer Ovarian stimulation outcomes are similar between transgender men and oocyte donation candidates, leading to a satisfactory mean number of cryopreserved oocytes. What is known already The reproductive potential of transgender people may be impaired by gender-affirming hormone treatment (GAHT) and is obviously suppressed in case of gender-affirming surgery involving bilateral ovariectomy and/or hysterectomy. The evolution of medical support for transgender people has made fertility preservation strategies possible. Fertility preservation in transgender men mainly relies on oocyte cryopreservation following controlled ovarian stimulation. However, few data are available to date on the subject, concerning small sample sizes, so no reliable conclusions can be drawn about the feasibility and efficiency of fertility preservation procedures in trans men. Study design, size, duration This retrospective study reports the results of fertility preservation counselling in 118 transgender men referred to our fertility centre from September 2018 to December 2022. Among them, 16 benefited from oocyte cryopreservation. Ovarian stimulation outcomes were compared with those of cisgender oocyte donors. Participants/materials, setting, methods Sixteen transgender men benefited from oocyte cryopreservation following ovarian stimulation and were matched 1:1 to cisgender oocyte donors according to age and body mass index. Antral follicle count and serum AMH levels were systematically measured. Primary outcomes included duration of the stimulation, total FSH dose, peak serum estradiol, oocyte yield, number of mature oocytes, and maturity rate (mature oocytes/total oocytes collected). Results were compared using Fisher’s exact or Wilcoxon’s rank sum tests. Main results and the role of chance One hundred and eighteen transgender men were referred in our centre for fertility preservation counselling. Among them, 95 asked for a medical consultation, and 86 came to the appointment. Following the consultation, only 16 (18.6%) finally decided to benefit from oocyte cryopreservation following ovarian stimulation, including 2 that had already started testosterone therapy. Sixteen presumably fertile oocyte donors were matched based on age and BMI. Mean age of trans’ men was 23.9 ± 5.0. Basal ovarian reserve tests showed satisfactory results in trans’ men and oocyte donors, with similar AMH (4.5 ± 2.3 vs. 4.4 ± 3.2, respectively, NS) but lower AFC in trans’men than in oocyte donors (13.8 ± 11.0 vs. 26.7 ± 10.1, p = 0.033). This result may be explained by a more frequent use of the abdominal approach to assess AFC in the transgender group. Ovarian stimulation outcomes were comparable, with no differences in the duration of stimulation, total FSH dose and peak estradiol levels, leading to a comparable mean number of mature oocyte (15.4 ± 9.9 vs. 17.2 ± 10.6, respectively, NS). Thirteen out 16 transgender men had 10 or more cryopreserved mature oocyte following a single procedure. Limitations, reasons for caution Only a low percentage of trans men that were referred to our centre finally chose to perform oocyte cryopreservation, leading to a small sample size. The majority of them had not started any GAHT, preventing us to evaluate the potential consequences of these treatments on ovarian stimulation outcomes. Wider implications of the findings While parenthood strategies for transgender people have long been ignored, fertility preservation is an important issue to consider, especially because medical treatments and surgeries may be undertaken in very young adults. Oocyte cryopreservation seems to represent a feasible way for trans men to preserve their fertility for future biological parenting. Trial registration number N/A

Outcomes of Different In Vitro Maturation Procedures for Oocyte Cryopreservation for Fertility Preservation and yet Another Live Birth in a Cancer Patient.

To ensure patient care in an oncological fertility preservation (FP) programme, specialists must provide technology that best suits the patients' clinical conditions. In vitro oocyte maturation (IVM) and ovarian tissue cryopreservation (OTC) are possible fertility preservation treatments for women in need of urgent oncological treatment. IVM consists of the retrieval of immature oocytes from small antral follicles, with no or minimal ovarian stimulation by gonadotropins. Therefore, IVM has become a pertinent option for fertility preservation, especially for cases whereby ovarian stimulation is unfeasible or contra-indicated. Existing data on immature oocytes, retrieved transvaginally (OPU-IVM) or extracted from ovarian tissue 'ex vivo' (OTO-IVM), are still limited on technical consistency, efficacy, and safety. The present retrospective cohort study includes 89 women undergoing fertility preservation using IVM methodologies and 26 women undergoing ovarian stimulation (OS) in concomitant period. In total, 533 immature oocytes were collected from IVM patients, achieving a maturation rate of 57% and 70% in OTO-IVM and 73% and 82% in OPU-IVM at 24 h and 48 h in culture, respectively. The observed high maturation rates might be due to the use of patients' serum in its innate status, i.e., without heat-inactivation. This permitted 7.6 ± 5.7 and 4.6 ± 4.9 oocytes to be vitrified in OTO-IVM and OPU-IVM, respectively, compared to 6.8 ± 4.6 from OS patients. Regarding OS patients, two of them underwent embryo transfer following the insemination of warmed oocytes after complete remission, resulting in a single live birth from one patient. Upon follow-up of two OTO-IVM patients after the termination of their oncological treatment, a total of 11 warmed oocytes lead to a transfer of a single embryo, but pregnancy was not achieved. From OPU-IVM, six embryos were transferred in three patients 4.25 years after oocyte vitrification, leading to the live birth of a healthy boy. The present case of live birth is among the first cases reported so far and supports the notion that IVM might be a relevant and safe FP option for cancer patients when oocyte preservation is required but ovarian stimulation is contra-indicated.

Expansion of the fertility preservation program to all newly diagnosed prepubertal oncology patients at a pediatric hospital.

10064 Background: As the majority of pediatric oncology patients survive their disease, generating a population of over 500,000 survivors in the United States, it is imperative to minimize the lifelong consequences of treatment, which includes temporary or permanent infertility caused by certain cancer treatments. Fertility preservation (FP), the opportunity to preserve one’s ability to have genetically-related children prior to damaging the reproductive system, is a young but rapidly expanding field and recent scientific advances have led to an expansion of options for prepubertal patients. A fertility consultation at diagnosis can provide patients and families with the opportunity to be informed regarding likelihood of gonadal dysfunction and to consider FP. Methods: After our pediatric hospital started to offer tissue cryopreservation, we initiated this evidence-based interventional quality improvement project. Our primary aim was to ensure that all newly diagnosed prepubertal oncology patients who met criteria for FP were correctly identified and offered an educational consultation and preservation. Inpatient admission lists and provider schedules were reviewed daily to identify patients. Treatment plans were discussed with providers to calculate infertility risk. All procedures were to be completed at the time of another procedure to increase likelihood of insurance coverage and minimize delays. Results: Between 07/15/22 and 11/30/22, 54 newly diagnosed prepubertal patients’ treatment plans were evaluated to determine treatment-related infertility risk using the Oncofertility Consortium Pediatric Initiative Network’s Risk Assessment tool. 15 patients were at a high level of significantly increased risk and were therefore eligible for consultation. 7 patients and their families received an educational consultation and 6 elected to undergo FP (3 TTCs; 3 OTCs). FP procedures did not cause a delay in starting treatment for any of these patients. 8 patients did not receive consultations. Reasons included the provider electing to proceed with treatment without consultation (4), the patient’s comorbidities (1), the inability to bundle FP with another procedure (1), and the family decision to decline consultation (1). Conclusions: Of the patients at high risk of infertility, only 46% received consultation with lack of referral prior to initiating treatment as the primary reason. Of the patients offered preservation methods, 86% elected to preserve their fertility, illustrating patient and family interest. Providers must acknowledge changes in practice and be informed on the impact that high-risk treatment protocols can have on fertility. An FP program with established policies processes can increase the likelihood that prepubertal patients at high risk for infertility are correctly identified, educated, and offered preservation.

Fertility Preservation in the Female Tumor Patients

As we all know, the prevalence of tumors is increasing, and with the development of medical examination and treatment methods, the detection rate and cure rate of tumors in young women are gradually increasing, and many patients still want to realize their dream of parenthood after realizing the basic survival need of "saving life". However, the progression of the disease itself and treatment methods such as radiotherapy and chemotherapy will cause irreversible damage to the fertility of patients, so it is particularly important for these patients to preserve fertility as soon as possible. Fertility preservation is in the ascendant in China, and tumor patients are one of its main indications. In recent years, the technology has made significant progress, the current mature technology includes ovarian tissue cryopreservation and transfer, egg and embryo freezing, etc., due to the lack of understanding and application of specific programs by doctors, many patients miss the opportunity of fertility preservation, in order to provide patients with more scientific fertility preservation programs, this paper combines the fertility preservation related research of female tumor patients at home and abroad in recent years, and reviews the current fertility preservation programs and progress of female tumor patients.

Gonadal development and function after immature testicular tissue banking as part of high-risk gonadotoxic treatment.

Experimental fertility preservation programs have been started to safeguard the future fertility of prepubertal and pubertal males requiring high-risk gonadotoxic treatment protocols. However, long-term follow-up studies evaluating the effects on their gonadal development and function related to the testicular biopsy procedure are rather limited. This two-center follow-up study (between 2002 and 2020) evaluated the gonadal development and function of a cohort of 59 prepubertal and pubertal males who have been offered immature testicular tissue banking (TTB) prior to conventional high-risk chemo- and/or radiotherapy (HR-C/R) or conditioning therapy before hematopoietic stem cell transplantation (CT-HSCT). The aim is to investigate the long-term impact of the testicular biopsy procedure and the high-risk gonadotoxic treatment. Testicular growth and the reproductive hormones luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), and inhibin B (INHB) were analyzed after treatment completion, and compared between males accepting TTB and those refusing TTB (control) as well as between HR-C/R and CT-HSCT treatment protocols. Of the 59 prepubertal and pubertal males included, 25 were treated by HR-C/R and 34 required CT-HSCT. TTB was accepted for 39 males and refused for 20 males. Most patients were prepubertal at diagnosis (85%), at TTB (79%), and at treatment completion (76%), and pubertal or postpubertal at their last follow-up visit (66%). After 5.0 (1.0-13.0) years post treatment, most patients show normal testicular volumes (83%) and normal LH (89%), FSH (87%), T (87%), and INHB (79%) serum levels. The testicular biopsy procedure did not have an effect on testicular growth, LH, FSH, T, and INHB. Significantly more small postpubertal testicular volumes (p=.0278) and low INHB serum levels (p=.0130) were recorded after CT-HSCT, especially after myeloablative conditioning. The clinical follow-up data demonstrate no effect related to the biopsy procedure, but a substantial risk for impaired gonadal development after high-risk gonadotoxic treatment, in particular myeloablative CT-HSCT. Longer follow-up studies with a larger study population are needed to confirm these preliminary findings.

MP43-11 EIGHT-YEAR FERTILITY PRESERVATION PROGRAM AT ATRIUM HEALTH WAKE FOREST BAPTIST FOCUSING ON SPERMATOGONIAL STEM CELL AND ROUND SPERMATID TECHNOLOGIES.

You have accessJournal of UrologyCME1 Apr 2023MP43-11 EIGHT-YEAR FERTILITY PRESERVATION PROGRAM AT ATRIUM HEALTH WAKE FOREST BAPTIST FOCUSING ON SPERMATOGONIAL STEM CELL AND ROUND SPERMATID TECHNOLOGIES. Adam Cohen, Janmejay Hingu, Nima Pourhabibi Zarandi, Guillermo Galdon, Omar Abdelaal, Banafsheh Nickmehr, Kimberly Stogner-Underwood, Stanley Kogan, Steve Hodges, Stuart Howards, Thomas McLean, Anthony Atala, and Hooman Sadri-Ardekani Adam CohenAdam Cohen More articles by this author , Janmejay HinguJanmejay Hingu More articles by this author , Nima Pourhabibi ZarandiNima Pourhabibi Zarandi More articles by this author , Guillermo GaldonGuillermo Galdon More articles by this author , Omar AbdelaalOmar Abdelaal More articles by this author , Banafsheh NickmehrBanafsheh Nickmehr More articles by this author , Kimberly Stogner-UnderwoodKimberly Stogner-Underwood More articles by this author , Stanley KoganStanley Kogan More articles by this author , Steve HodgesSteve Hodges More articles by this author , Stuart HowardsStuart Howards More articles by this author , Thomas McLeanThomas McLean More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and Hooman Sadri-ArdekaniHooman Sadri-Ardekani More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003289.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Male infertility has been associated with many disease processes, including chemotherapy and radiotherapy for cancer, undescended testis (UDT), and genetic conditions such as Klinefelter Syndrome (KS). Preservation of human spermatogonial stem cells (SSC) may be a suitable option for young male patients at risk of losing fertility. With optimal cryopreservation, cell viability can be retained to reestablish spermatogenesis in the future through in vitro germ cell differentiation or round spermatid injection. METHODS: Urology and hematology-oncology patients at Atrium Health Wake Forest Baptist with malignancy, UDT, or KS were approached for consent and assent (if ≥seven years old) for the fertility preservation program. If participation was accepted, patients were given the option to undergo a fertility-preserving testicular biopsy, preferably during general anesthesia for other procedures such as Port-a-Cath placement, orchidopexy, or lymph node biopsy. They were also allowed to enroll as a registry participant (non-biopsy control). Histological and microbiological analyses were performed on biopsies before being cryopreserved. Further data were obtained retrospectively via chart review. RESULTS: Over eight years of banking, a total of 202 patients were approached for the study, with 125 (61.8%) undergoing a biopsy, 39 (19.3%) enrolling as non-biopsy control, and 38 (18.8%) declining participation. The mean age at which biopsy was performed across all diagnoses was 8.39 years (range 0.61 to 37.39). Of the 125 biopsy patients, 73% opted to donate a portion (up to 20%) of the tissue for basic research with no direct benefit to the donor. Of the biopsied patients, 76 (60.8%) were performed for cancer, 38 (30.4%) for unilateral or bilateral UDT, and 11 (8.8%) for KS. Based on PRM1 immunofluorescence, 48.3% of specimens had round spermatids. Short-term complications for testicular biopsy, primarily pain and swelling, were typically mild to moderate. CONCLUSIONS: A multidisciplinary approach can accommodate patients at risk for reduced future fertility due to various pathologies. The preservation of testicular tissue at present can aid in future fertility restoration through techniques such as in vitro SSC differentiation or round cell spermatid injection (ROSI). Source of Funding: N/A © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e605 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Adam Cohen More articles by this author Janmejay Hingu More articles by this author Nima Pourhabibi Zarandi More articles by this author Guillermo Galdon More articles by this author Omar Abdelaal More articles by this author Banafsheh Nickmehr More articles by this author Kimberly Stogner-Underwood More articles by this author Stanley Kogan More articles by this author Steve Hodges More articles by this author Stuart Howards More articles by this author Thomas McLean More articles by this author Anthony Atala More articles by this author Hooman Sadri-Ardekani More articles by this author Expand All Advertisement PDF downloadLoading ...

Long-Term Fertility Function Sequelae in Young Male Cancer Survivors.

With advances in cancer treatment, such as cytotoxic chemotherapy and radiotherapy, grave new sequelae of treatment have emerged for young cancer survivors. One sequela that cannot be overlooked is male infertility, with reportedly 15% to 30% of cancer survivors losing their fertility potential. Cytotoxic therapy influences spermatogenesis at least temporarily, and in some cases, permanently. The degree of spermatogenesis impairment depends on the combination of drugs used, their cumulative dose, and the level of radiation. The American Society of Clinical Oncology has created an index to classify the risks to fertility based on treatment. Medical professionals currently use this risk classification in fertility preservation (FP) programs. FP programs are currently being promoted to prevent spermatogenesis failure resulting from cancer treatment. For patients who are able to ejaculate and whose semen contains sperm, the semen (sperm) is cryopreserved. Moreover, for patients who lack the ability to ejaculate, those with azoospermia or severe oligozoospermia, and those who have not attained puberty (i.e., spermatogenesis has not begun), testicular biopsy is performed to collect the sperm or germ cells and cryopreserve them. This method of culturing germ cells to differentiate the sperm has been successful in some animal models, but not in humans. FP has recently gained popularity; however, some oncologists and medical professionals involved in cancer treatment still lack adequate knowledge of these procedures. This hinders the dissemination of information to patients and the execution of FP. Information sharing and collaboration between reproductive medicine specialists and oncologists is extremely important for the development of FP. In Japan, the network of clinics and hospitals that support FP is expanding across prefectures.