Human reproduction open最新文献

{"title":"Heterogeneity in semen-mediated tolerogenic responses: defining immunological determinants of reproductive compatibility.","authors":"Shahrokh Paktinat, Michael G Gravett, Sean M Hughes, Abigail L P Spray, Germán G Gornalusse, Florian Hladik, Lucia Vojtech","doi":"10.1093/hropen/hoag064","DOIUrl":"10.1093/hropen/hoag064","url":null,"abstract":"<p><strong>Study question: </strong>Do particular pairs of individuals modeling sexual pairs exhibit differences in the induction of semen-mediated long-lasting tolerance in support of pregnancy?</p><p><strong>Summary answer: </strong>Yes, factors from both sexes contribute to the variability in the induction of long-lived regulatory T cells (Tregs) in response to semen, with factors related to the recipient seeming to slightly outweigh the factors related to the semen donor.</p><p><strong>What is known already: </strong>Semen contains multiple immunomodulatory factors that can influence reproductive tract immunity in the recipient, promote tolerance to antigens from the semen donor (designated henceforth as 'paternal'), and ultimately support successful pregnancy. These factors include the soluble fraction containing prostaglandins and cytokines, as well as a very high concentration of extracellular vesicles (EVs). Both soluble and vesicular fractions of semen have been shown to promote tolerogenic phenotypes and migration in antigen-presenting cells (APCs), and to induce Tregs. This study was undertaken to understand the variation in tolerance induction between pairs of individuals modeling sexually active couples.</p><p><strong>Study design size duration: </strong>We designed an <i>in vitro</i> pairing study using random semen donors (N = 11) and peripheral mononuclear cells (PBMCs) from random self-identified female recipients (N = 3). Semen was fractionated into vesicle-enriched (semen extracellular vesicles; SEVs) and soluble (vesicle-depleted semen plasma; VDSP) fractions. Monocyte-derived dendritic cells (MoDCs) from each of the recipients were exposed to semen fractions from each donor, then cocultured with autologous naïve T cells to assess tolerogenic responses, specifically, generation of Tregs (FOXP3+ CD25Hi CD127Lo CD4+ T cells) and their phenotypic differences (TIGIT+, PD-1+ CTLA-4+, or AREG+ Tregs). These cells were then screened for T cell receptor-dependent activation by activation-induced marker (AIM) expression assay. These responses were compared across all pairs of semen donors and recipient cells. In addition, semen fractions were screened for candidate mediators of tolerance, such as human leukocyte antigen G (HLA-G) and adenosine-generating enzymes.</p><p><strong>Participants/materials setting methods: </strong>This study used an <i>in vitro</i> human cell culture approach in which primary blood-derived immune cells were exposed to semen-derived vesicular and non-vesicular fractions. Blood samples were obtained from healthy self-identified female donors, and semen samples were obtained from healthy participants who provided initial screening specimens for a contraceptive study. Semen-derived vesicular and non-vesicular fractions were characterized by western-blot analysis for markers associated with immune tolerance. Primary immune cells were exposed to these semen fractions, and the major endpoints included changes in Tr","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag064"},"PeriodicalIF":8.6,"publicationDate":"2026-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13446728/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148686898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An artificial intelligence real-time rare sperm detection system for intraoperative microsurgical testicular sperm extraction.","authors":"Sha Han, Shuai Xu, Haojun Hu, Zijue Zhu, Furong Bai, Cunzhong Deng, Yuhua Huang, Chenwang Zhang, Yifan Sun, Haowei Bai, Chencheng Yao, Fujun Zhao, Zheng Li, Dengyang Zhao, Peng Li","doi":"10.1093/hropen/hoag062","DOIUrl":"10.1093/hropen/hoag062","url":null,"abstract":"<p><strong>Study question: </strong>How can rare spermatozoa be identified efficiently during microsurgical testicular sperm extraction (micro-TESE)?</p><p><strong>Summary answer: </strong>An artificial intelligence (AI)-assisted system was developed to flag candidate rare spermatozoa in real time during micro-TESE, and it may support embryologists as a decision-support tool.</p><p><strong>What is known already: </strong>Patients with non-obstructive azoospermia (NOA) can obtain sperm for procreation through micro-TESE. During this procedure, sperm retrieval primarily relies on embryologists or laboratory technicians visually searching for sperm under a microscope, which is not only laborious and inherently subjective but also susceptible to errors. Although AI technology has been applied to identify trace amounts of sperm, existing models lack sufficient efficiency and true real-time performance.</p><p><strong>Study design size duration: </strong>This study included model development followed by a single-centre clinical evaluation. An improved YOLO (You Only Look Once)-based rare sperm detection model, termed YOLOv11-RSD, was developed using microscopy data from 1165 surgical patients, comprising 1932 image samples containing a total of 5032 annotated sperm objects with confirmed identification. Clinical evaluation was performed between May 2024 and July 2025. Performance was assessed across confidence thresholds in obstructive azoospermia (OA) patients with normal spermatogenesis, and the system was then applied during micro-TESE in NOA patients and compared with routine embryologist assessment.</p><p><strong>Participants/materials setting methods: </strong>The model was developed using testicular sperm microscopy images collected at a single hospital. Real-time clinical feasibility was evaluated in 10 OA cases and 30 NOA cases. Embryologist assessment was used as the reference standard, and performance was assessed using PPV, sensitivity, <i>F</i>1-score, and 95% confidence intervals. Discordant AI-assisted detections were reviewed by embryologists in real time.</p><p><strong>Main results and the role of chance: </strong>YOLOv11-RSD achieved real-time detection of candidate spermatozoa in microscopy images with high sensitivity and acceptable PPV under the selected operating threshold. Compared with baseline YOLOv11, YOLOv11-RSD showed improved overall detection performance across representative evaluation settings. In OA cases, the system achieved high sensitivity for sperm detection, reaching up to 96.7% across evaluated thresholds. During micro-TESE in NOA patients, at a confidence threshold of 0.50, positive predictive value (PPV), sensitivity, and <i>F</i>1-score were 80.58%, 96.11%, and 87.66%, respectively. The system highlighted candidate spermatozoa that were not identified during the initial manual assessment in six NOA cases, including two cases initially classified as sperm-negative; these findings were confirmed upon immediate re-","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag062"},"PeriodicalIF":8.6,"publicationDate":"2026-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13437015/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148674416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Don't panic: IVF efficiency in the USA remains strong.","authors":"Micah J Hill, Jennifer F Kawwass, Jennifer L Eaton","doi":"10.1093/hropen/hoag060","DOIUrl":"10.1093/hropen/hoag060","url":null,"abstract":"","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag060"},"PeriodicalIF":8.6,"publicationDate":"2026-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13428317/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148654842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to: Telehealth cognitive behavioural therapy improves health-related quality of life and pain in endometriosis: the Healing Pelvic Pain Intervention (HaPPI)-a randomized controlled trial.","authors":"","doi":"10.1093/hropen/hoag059","DOIUrl":"https://doi.org/10.1093/hropen/hoag059","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.1093/hropen/hoag006.].</p>","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag059"},"PeriodicalIF":8.6,"publicationDate":"2026-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13310949/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148354964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Don't panic? Or don't conflate?","authors":"Norbert Gleicher, Sonia Gayete-Lafuente, Lara Guijarro-Baude, Pasquale Patrizio, David F Albertini, David H Barad","doi":"10.1093/hropen/hoag061","DOIUrl":"10.1093/hropen/hoag061","url":null,"abstract":"","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag061"},"PeriodicalIF":8.6,"publicationDate":"2026-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13428318/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148654910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Beyond cardiovascular risk: is 'normal' blood pressure too high for reproductive success?","authors":"Berthold Hocher, Shujuan Ma","doi":"10.1093/hropen/hoag057","DOIUrl":"10.1093/hropen/hoag057","url":null,"abstract":"<p><p>The relationship between blood pressure (BP) and reproductive health is increasingly recognized as a clinically relevant and biologically plausible axis influencing fertility outcomes. Evidence from ART cohorts suggests that reproductive success may decline across a continuum of increasing BP, extending below conventional hypertensive thresholds. Observational studies have linked male prehypertension with impaired semen quality and reduced ART success, and maternal prepregnancy systolic BP with lower live birth rates, higher miscarriage risk, and a greater burden of hypertensive pregnancy complications. These findings raise a provocative question: are BP levels considered acceptable for long-term cardiovascular prevention also optimal for implantation, placentation, and live birth? This Opinion article argues that preconception BP should be considered a potentially informative reproductive vascular marker, while clearly distinguishing BP as a possible causal determinant from BP as a surrogate of underlying vascular-metabolic health. Potential mechanisms include endothelial dysfunction, impaired nitric oxide signalling, reduced uterine perfusion, vascular inflammation, and altered remodelling affecting both the maternal environment and gamete quality. At the same time, causality remains unproven because current evidence is predominantly observational and vulnerable to residual confounding. A focused research agenda is therefore needed to test whether preconception BP optimization can improve reproductive outcomes and to determine whether reproductive medicine requires endpoint-specific BP thresholds.</p>","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag057"},"PeriodicalIF":8.6,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13353218/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148426922","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>All-in-one</i> exome sequencing approach for genetic testing of unexplained premature ovarian insufficiency.","authors":"Anu Valkna, Triin Kikas, Ülle Jakovlev, Oliver Mõttus, Avirup Dutta, Külli Erlang, Margus Punab, Kristiina Rull, Maris Laan","doi":"10.1093/hropen/hoag058","DOIUrl":"10.1093/hropen/hoag058","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Study question: &lt;/strong&gt;Is exome sequencing (ES) an efficient approach for simultaneous analysis of causative single gene defects and copy number variants (CNV) in unexplained premature ovarian insufficiency (POI)?&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Summary answer: &lt;/strong&gt;Among 51 idiopathic POI cases, a conservative molecular diagnostic yield of 12% was achieved with equal contribution of pathogenic or likely pathogenic (P/LP) monogenic causes and pathogenic microdeletions.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;What is known already: &lt;/strong&gt;POI is a clinically and genetically heterogeneous condition, yet most cases remain idiopathic. Although numerous monogenic causes and CNVs have been implicated in POI, standardized clinical guidelines for comprehensive genetic analysis are still lacking.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Study design size duration: &lt;/strong&gt;This observational study of 51 unexplained POI cases implemented an &lt;i&gt;all-in-one&lt;/i&gt; ES-based approach to analyze P and LP variants in 288 POI candidate genes and large (&gt;0.5 Mb) pathogenic CNVs leading to POI.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Participants/materials setting methods: &lt;/strong&gt;Patients were recruited and phenotyped at two tertiary care women's health and endocrinology centers in Estonia. Bioinformatic processing of ES data and assessment of rare causal variants in POI were performed using an in-house analysis pipeline including automated filtering and manual assessment for pathogenicity, followed by experimental validation via Sanger sequencing (monogenic findings) and chromosomal microarray analysis (CNVs).&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Main results and the role of chance: &lt;/strong&gt;Six of 51 unexplained POI cases had confident molecular findings, including three cases with P/LP monogenic variants and three carrying large pathogenic CNVs. Overall molecular diagnostic yield was estimated to be 12% (6 of 51). There was a statistically significant overrepresentation of likely causal genetic findings in primary amenorrhea (5 of 9 cases, 56%) compared to secondary amenorrhea/oligomenorrhea (1 of 42 cases, 2%) (&lt;i&gt;P &lt;/i&gt;= 3.0 × 10&lt;sup&gt;-4&lt;/sup&gt;). Primary amenorrhea cases presented diverse findings-TP63 p.(Arg97Gly) in three sisters (including one identified by cascade screening), 15q25.2 microdeletion in two unrelated subjects (POI relevant haploinsufficient genes: &lt;i&gt;CPEB1, BNC1&lt;/i&gt;), and NR5A1 p.(Gly328Arg) in one case. A secondary amenorrhea case with ∼8.5 Mb microdeletion at Xq27.3-Xq28 enabled us to narrow down a critical region for non-recurrent Xq terminal losses causing POI (relevant haploinsufficient genes &lt;i&gt;FMR1&lt;/i&gt;, &lt;i&gt;AFF2&lt;/i&gt;, &lt;i&gt;CETN2&lt;/i&gt;, &lt;i&gt;HAUS7&lt;/i&gt;, and &lt;i&gt;EMD&lt;/i&gt;). Additionally, heterozygous variants &lt;i&gt;TBX6&lt;/i&gt; c.622-2A&gt;T, &lt;i&gt;EXO1&lt;/i&gt; c.136del, and NR2F2 p.(Val307Ala), as well as 1q21.1 microdeletion (two carriers), 1p36.22, and 12q21.1 microduplications were classified as potentially interesting Variants of Uncertain Significance (VUS) that require validation before their causal link to isolated POI can be assigned. &lt;i&gt;TBX6&lt;/i&gt; c.622-2A&gt;T an","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag058"},"PeriodicalIF":8.6,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13372669/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148474668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Predicting natural conception leading to live birth for couples with infertility: a single-centre population-based cohort study of 7086 couples.","authors":"Natalie J Cameron, Kate Brian, David J McLernon, Siladitya Bhattacharya","doi":"10.1093/hropen/hoag056","DOIUrl":"10.1093/hropen/hoag056","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Study question: &lt;/strong&gt;Can a model be developed to predict natural conception leading to live birth over a period of 12 months in couples with different causes of infertility?&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Summary answer: &lt;/strong&gt;We developed and validated a novel clinical prediction model that can be used to provide individualized estimates of the chance of a natural conception (leading to live birth) over a 12-month period for couples with different causes of infertility, including tubal factor, anovulation, male factor, unexplained, other (cervical, uterine, or sexual factor), and endometriosis.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;What is known already: &lt;/strong&gt;Existing prediction models for natural conception are primarily aimed at infertile couples with no identified cause (unexplained), mild male factor, or minimal endometriosis. Currently, there is no available model that is applicable across a wide range of fertility diagnoses at the point of initial assessment.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Study design size duration: &lt;/strong&gt;A population-based cohort study based on data from a single large tertiary UK fertility centre serving the entirety of the North of Scotland, including all 9757 couples who registered at the clinic for the first time from 1998 to 2015.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Participants/materials setting methods: &lt;/strong&gt;Using a Cox proportional hazards survival model, we estimated the chance of conception within the first year from diagnosis of infertility leading to live birth. The predictive accuracy of the model was assessed using discrimination and calibration measures in an internal validation. The clinical utility of the model was assessed using decision curve analysis.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Main results and the role of chance: &lt;/strong&gt;After exclusions, 7086 couples with infertility were included, of whom 891 (13%) had a natural conception within 1 year of diagnosis leading to live birth. Longer duration of infertility [hazard ratio (HR): 0.64 (95% CI: 0.57-0.72)], increasing female age [0.62 (0.55-0.70)], and tubal infertility [0.62 (0.47-0.81)] had the strongest influence on reducing the chance of natural conception within 1 year leading to live birth. Conversely, previous pregnancy [1.30 (1.12-1.50)] and unexplained infertility [1.29 (1.00-1.67)] were most strongly associated with increased chances of live birth. For example, a 25-year-old woman with 1 year of secondary unexplained infertility has a 33.8% predicted chance of having a baby resulting from natural conception within a year of diagnosis. In contrast, a 35-year-old woman with tubal infertility and 1 year of primary infertility has an estimated 6.0% predicted chance of live birth over a similar time horizon. The model demonstrated acceptable discrimination (optimism-corrected C-statistic 0.645) and good calibration in internal validation. Decision curve analysis demonstrated that applying the model at a 20% probability threshold would correctly identify an additional 7.2 couples per 100 as being likely to benefit ","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag056"},"PeriodicalIF":8.6,"publicationDate":"2026-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13353215/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148426955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Should we routinely perform sperm DNA fragmentation testing in fertility clinics?","authors":"Srisailesh Vitthala, Tarek El-Toukhy, Abha Maheshwari","doi":"10.1093/hropen/hoag047","DOIUrl":"10.1093/hropen/hoag047","url":null,"abstract":"<p><p>Routine semen analysis provides limited diagnostic and prognostic insight into male reproductive potential, contributing to the increasing use of sperm DNA fragmentation (SDF) testing in fertility evaluation. However, the clinical application of SDF testing appears to outpace the strength and consistency of the supporting evidence. This opinion paper critically appraises SDF testing through established screening, diagnostic, and prognostic test-performance frameworks, drawing on published observational studies, meta-analyses, and international guideline statements across natural conception and ART. The available evidence indicates that SDF testing does not meet the criteria for population-level screening or for routine diagnostic or prognostic use in unselected infertile populations and is frequently applied beyond its validated scope. Interpretation is further limited by assay heterogeneity, lack of standardization, variable thresholds, reliance on surrogate outcomes, and the limited effectiveness of available interventions. Routine implementation therefore risks overdiagnosis, unnecessary interventions, and premature escalation to assisted reproduction without proven benefit. When applied selectively within an aetiology-driven framework, particularly in defined subgroups, SDF testing may provide adjunctive risk stratification rather than deterministic clinical guidance. All stakeholders must be made aware of these limitations before the test is offered, interpreted, or used to modify treatment.</p>","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag047"},"PeriodicalIF":8.6,"publicationDate":"2026-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13274538/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148280282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human iPSCs derived from cryopreserved testicular somatic cells enable germline regeneration in childhood cancer survivors.","authors":"Tiago Macedo, Claudia De Guidi, Leah Nic Aodha, Nageswara Rao Boggavarapu, Maja Piechocka, Xuan Ye, Francesca Mastropasqua, Victoria Keros, Ulrika Norén Nyström, Per Frisk, Pia Johansson, Kristiina Tammimies, Yoni Baert, Kirsi Jahnukainen, Jan-Bernd Stukenborg, João Pedro Alves-Lopes","doi":"10.1093/hropen/hoag054","DOIUrl":"10.1093/hropen/hoag054","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Study question: &lt;/strong&gt;Can cryopreserved primary testicular somatic cells from childhood cancer survivors with severely decreased fertility potential be reprogrammed into human-induced pluripotent stem cells (hiPSCs) competent for efficient specification into early human germ cells?&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Summary answer: &lt;/strong&gt;Primary testicular somatic cells from cryopreserved testicular samples with severely compromised spermatogonial pools can be reprogrammed into hiPSCs using a non-genome-integrating, feeder-free approach and subsequently differentiated into human primordial germ cell-like cells (hPGCLCs) with high efficiency.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;What is known already: &lt;/strong&gt;Infertility is one of the most concerning long-term side effects of cancer therapy in prepubertal boys, yet it remains largely unaddressed. Worldwide, biobanks storing cryopreserved immature testicular tissue are expanding to support the development of fertility preservation strategies, including future tissue and cell transplantation approaches. However, whether these samples can also serve as starting material to generate hiPSCs and subsequently differentiate into &lt;i&gt;in vitro&lt;/i&gt;-derived germ cells remains largely unexplored. Currently, there are established protocols for non-genome-integrating reprogramming of somatic cells into hiPSCs and for hPGCLC specification that could be applied to cryopreserved testicular tissue, with special relevance for patient samples severely depleted of germ cells.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Study design size duration: &lt;/strong&gt;Two biological replicates of cryopreserved prepubertal testicular tissue were used to obtain primary somatic cells, which were reprogrammed into hiPSCs and subsequently differentiated into hPGCLCs. This experimental pipeline had a duration of approximately four months.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Participants/materials setting methods: &lt;/strong&gt;Cryopreserved testicular tissue samples were obtained from two prepubertal cancer patients (6.2 and 6.3 years old) with depleted spermatogonial pools (spermatogonia count per round tubular cross-section of 0.04 and 0.02, and age-standardized &lt;i&gt;Z&lt;/i&gt;-scores of -17.62 and -20.48). These samples were subsequently used to derive primary testicular somatic cell cultures, which were then reprogrammed into hiPSCs using a clinically compatible, non-genome-integrating mRNA-based method under feeder-free conditions. The resulting hiPSC lines were validated and subsequently differentiated into hPGCLCs using two different specification protocols. The transcriptomic profiles of the hiPSCs and their derived hPGCLCs were verified using bulk RNA sequencing.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Main results and the role of chance: &lt;/strong&gt;Here, we present the first successful generation of hiPSCs from cryopreserved testicular somatic cells of childhood cancer patients with a severely depleted germ cell pool. We accomplished this using a non-genome-integrating mRNA-based reprogramming approach. We further demonstrated the specification","PeriodicalId":73264,"journal":{"name":"Human reproduction open","volume":"2026 3","pages":"hoag054"},"PeriodicalIF":8.6,"publicationDate":"2026-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13322296/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148371270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}