BACKGROUND:Cryogenic Dewars used for IVF cryostorage are commonly evaluated through periodic inspections and evaporation measurements, which may delay the detection of insulation degradation or lead to premature Dewar retirement. Conventional failure models emphasize catastrophic vacuum loss; however, field observations suggest that many Dewars exhibit transient or progressive thermal anomalies prior to permanent failure. METHODS:A retrospective observational study was conducted using longitudinal liquid nitrogen (LN2) temperature telemetry data acquired from static IVF cryostorage units operating under continuous clinical conditions. Existing temperature data were evaluated to identify transient deviations, recovery behavior, event frequency, and long-term baseline drift. These features were integrated into a composite Dewar Health Index (DHI) designed to characterize insulation performance and degradation risk over time without direct measurement of vacuum pressure. In parallel, the normal evaporation rate, the steady-state rate of LN2 evaporative loss derived from continuous level telemetry, was evaluated as an independent, energy-based measure of heat leak into the cryogenic system. Trend-based analytics were applied to distinguish temporary disturbances from progressive and permanent degradation modes. Analyses were performed on randomly sampled, de-identified telemetry drawn from an extensive cryostorage monitoring database comprising over 1000 active zones with continuous 5-minute data acquisition. RESULTS:Cryostorage Dewars exhibited nonbinary thermal behavior, including transient temperature deviations followed by recovery, consistent with known cryogenic phenomena and dynamics. Dewars exhibiting increasing excursion frequency, prolonged recovery times, and sustained baseline temperature drift showed patterns consistent with progressive degradation prior to retirement. The DHI differentiated stable, degrading, and high-risk Dewars solely based on longitudinal temperature telemetry. CONCLUSIONS:Longitudinal temperature analytics provide a noninvasive means of assessing the health of cryogenic Dewars in IVF cryostorage applications. The DHI enables early identification of degradation trends and supports risk stratification and maintenance planning without changes to clinical workflows. These findings highlight the value of continuous monitoring and data-driven assessment for improving cryostorage reliability and safety in assisted reproductive technology environments.
The terminology "Biobank" is used for the organized collection of biological materials consisting of tissue samples, blood, serum, body fluids, and DNA/RNA materials for scientific research. A biobank extracts the information from the biospecimens to generate databases, which are continuously updated. Over the years, there have been significant advances in digital pathology and artificial intelligence (AI). Digital pathology primarily relies on whole-slide imaging and cloud computing. AI, on the contrary, is able to handle vast amount of data and to recognize the patterns of the histopathology images in more meaningful ways. A digital biobank is defined as the integration of the bio-specimens with whole-slide images, molecular data, longitudinal clinical information, and metadata. Digital biobanking can do real-time monitoring of the quality of the biospecimen, rapid retrieval of data, and AI-based research work. It may take a pivotal role in precision medicine by utilizing AI-based models for patient management. This review discusses the evolution from conventional to digital biobanking and emphasizes AI-based quality control, multimodal data integration, and predictive analysis in oncology. In addition, virtual biobanks, hybrid physical-virtual models, and digital twins are also discussed. The article also discusses the key challenges related to the implementation of digital biobanking, including standardization, interoperability, and sustainability.
BACKGROUND:This study compared the isolation of peripheral blood mononuclear cells (PBMCs) using two methods. The first method was our routine Ficoll-Paque density gradient centrifugation isolation, which had been in place for many years. The second method was a new kit produced by Miltenyi on an autoMACS® Pro Separator with the new human Whole Blood PBMC Isolation Kit. The goal was to determine the potential to replace the manual Ficoll-Paque method with this new semi-automated method to improve standardization and efficiency of PBMC isolation, a pivotal step in processing whole blood cells for our biobanking purposes. METHODS:PBMCs were isolated from the peripheral blood of 10 healthy donors by routine Ficoll-Paque density gradient centrifugation and an autoMACS® Pro Separator by Miltenyi. Isolated PBMCs were analyzed by flow cytometry, and the proportions of lymphocyte and monocyte subtypes were determined. In addition, a panel of 16 kinase inhibitors was used to determine differences in drug responses using an assay that determines cell death over time. RESULTS AND CONCLUSION:Compared with Ficoll-Paque, the Miltenyi method yielded approximately 20 percent lower PBMC yield with no significant difference in cell viability and was superior when removing red blood cells. Proportions of some subpopulations (total T cells, B cells, and classical and intermediate monocytes) were altered based on the isolation method. Significant differences in drug responses were observed between the two methods. Moreover, the Miltenyi method introduced additional variability in drug sensitivity assays. These findings demonstrate that the two methods are not equivalent, which may limit the extent to which the Miltenyi method can improve standardization and reduce laboratory technician time in the isolation of PBMCs for biobanking.
AIM:Japanese quail offers massive potential as a source of animal protein and could fulfil the global surge in the demand for poultry meat. Sperm cryopreservation and assisted reproduction are pivotal in its propagation, as they simultaneously address various challenges like inbreeding, unreceptiveness towards new mates and unmanaged larger flocks. However, freezing procedures promote oxidative stress, disturb the redox homeostasis, and deteriorate sperm quality. Therefore, the current experiment was planned to evaluate the addition of catalase (CAT) antioxidant in the extender for Japanese quail sperm quality, mitochondrial activity, free radical scavenging activity and lipid peroxidation (LPO) during cryopreservation. MATERIALS AND METHODS:Semen was collected from 30 quails, pooled, divided and diluted with NaCl extender (115) containing 10 U CAT/mL, 30 U CAT/mL, 50 U CAT/mL and control. Extended semen (37°C) was gradually cooled to 20°C, then equilibrated at 4°C with 1% dimethylacetamide (DMA) and cryopreserved in liquid nitrogen. The sperm quality and biochemical parameters were assessed at the post-dilution, post-cooling, post-equilibration and post-thaw (0-hour and half-hour) stages of cryopreservation. RESULTS:The sperm motility, viability, plasma membrane and acrosomal integrity, mitochondrial activity and free radical scavenging activity were documented as highest (p < 0.05) with 10 U CAT/mL compared to other concentrations and the control at all the stages of cryopreservation. The LPO in sperm and seminal plasma was recorded lowest (p < 0.05) with 10 U CAT/mL at all the freezing stages. Moreover, at the post-thaw half-hour stage, only 10 U CAT/mL maintained the sperm quality. CONCLUSION:The addition of 10 U CAT/mL efficiently preserved and protected the sperm plasma membrane, acrosome, and mitochondrial activity as an antioxidative agent against the free radicals produced at chilling temperatures.
Introduction: The NIH-funded initiative Developmental Genotype-Tissue Expression (dGTEx) is an ongoing large resource of nonhuman primate (NHP) samples in which multiple organs are being collected, subdissected and preserved to build a unique tissue biorepository. Primates share close evolutionary relationships with humans, allowing for strong translation of research findings to human health. Objective: Generate a resource consisting of the two most frequently used NHP species in research the rhesus macaque ( Macaca mulatta ), which is widely available and has a well-studied genome, and the common marmoset ( Callithrix jacchus ), which is favored for studies of brain development and neurodegenerative disease. Methods: The dGTEx project is building (1) a genetic repository that will provide genome and tissue-based RNA sequencing for these Old and New World primate species to identify how genetic variations affect normal gene expression and to understand mechanisms of disease, and (2) an expansive histopathology resource of preserved individual organs. Detailed animal history, demographics, and sample metadata are documented in a secure database. Samples are being obtained from first (rhesus only), second, and third trimester fetal cohorts (with accompanying gestational tissues) as well as neonatal, infant, juvenile, and adult animal timepoints balanced for male and female to facilitate sex-specific data analysis. Results: We detail the sampling sites, preservation methods, and accompanying histology review that allows for confirmation of tissue type and quality, in addition to noting any pathological findings. Conclusion: This comprehensive NHP resource enables developmental investigations across the lifespan, from prenatal and postnatal stages through to adulthood.
Tumor-derived model biobanks comprise patient-derived xenografts, organoids, cell models or cultures, and selected paired derivatives linked to clinical, pathological, molecular, and functional data. In China, patient-derived organoid and xenograft resources have expanded through clinical and academic studies, enterprise-operated platforms, and conventional hospital biobank support for clinical sourcing and quality management. Published studies demonstrate disease-focused living resources in common tumors as well as selected premalignant and less common settings. Clinical and academic groups provide clinically annotated specimens and outcome context, whereas enterprise-operated platforms describe model-generation workflows, functional testing modules, database interfaces, and preclinical service capacity. Together, these activities suggest a transition from project-based model collections toward translational research infrastructure. However, public catalogues, model-level quality descriptors, case-model-data linkage, and access mechanisms remain uneven. This review summarizes representative Chinese resources and enterprise platform types, discusses international benchmark functions for model-biobank maturity, and emphasizes quality transparency for external verification and resource discovery. Future development should strengthen minimum information fields, quality reporting, and governance mechanisms that allow model resources to be discovered and used under appropriate access control.
INTRODUCTION:The gene expression patterns that drive human development are not fully understood. OBJECTIVES:To address this major public health need for pediatric clinicians, scientists, parents, and children, the National Institute of Health established the Developmental Genotype-Tissue Expression (dGTEx) project to study tissue-specific gene expression during the normal pediatric development of human organs, tissues, and cells. METHODS:Obtaining suitable, well-annotated pediatric biospecimens for innovative molecular methodologies and rigorous data analysis is both a high priority and unmet health need for the pediatric research community. RESULTS:Here, we report on the advancements and challenges in dGTEx with pediatric-specific tissue collection methodologies, metadata collection, and tissue verification processes. CONCLUSION:Collectively, the procedures, data, and biospecimens from dGTEx will provide an unparalleled resource for pediatric clinicians and scientists. The aim of establishing this resource is to gain a better understanding of pediatric gene expression and regulation across all body systems and stages of development, with potentially deeper insights into, and the development of new treatment options for, both diseases of childhood and later-onset, developmental-linked diseases in adults.
INTRODUCTION:Cryopreservation of single cells is well established; however, complex tissues remain challenging due to cellular heterogeneity and ice crystal-induced damage. Alterations in tissue architecture, for example, ice crystal formation, can compromise diagnostic usability, especially when subtle ultrastructural features are essential for interpretation. OBJECTIVES:To assess the impact of different freezing protocols, with or without cryoprotective agents (CPAs), on tissue (ultra)structure and nucleic acid integrity. METHODS:Setup A: murine livers were preserved using combinations of permeating (glycerol, 3-O-methyl-d-glucose) and nonpermeating CPAs (trehalose, polyethylene glycol [PEG]), then snap frozen in vapor phase of liquid nitrogen (VPLN, -196°C). Ultrastructure was analyzed by electron microscopy (EM). Setup B: comparison of freezing protocols (snap freezing in VPLN, controlled-rate freezing to -80°C, direct freezing at -80°C and -20°C) and prefreezing delays (0, 60, 90 minutes) with and without CPAs. EM, RNA/DNA integrity, and hematoxylin and eosin-stained cryosections were evaluated. RESULTS:All freezing protocols revealed nuclear changes, mitochondrial damage, and cytoplasmic abnormalities (Setup A/B). PEG-based CPA combinations were associated with milder ultrastructural alterations compared with glycerol-based variants and were selected for setup B. Controlled-rate and direct freezing at -80°C induced cytoplasmic changes. Delayed freezing increased mitochondrial irregularities; CPAs offered partial protection at 60 minutes but not at 90 minutes delayed freezing. Light microscopy and nucleic acid integrity showed no apparent differences. CONCLUSION:PEG-based CPAs partially stabilized ultrastructure, but prefreezing delay appeared to be the dominant factor influencing tissue quality. Rapid processing and optimized freezing protocols are essential to minimize cryodamage. Combined (ultra)structural and molecular evaluation is recommended. Results should be interpreted in the context of freezing method, particularly for ultrastructural analyses.
Standardized biospecimen banking is foundational for biomedical research and eventually underpins population health outcomes. In many low- and middle-income countries (LMICs) biobanking remains fragmented and weakly governed, limiting both scientific value and public trust in the ensuing research. This commentary presents insights from a stakeholder dialogue at a national pathology conference in Pakistan. The discourse identified drivers of human biospecimen banking fragmentation and governance gaps in the country, while mapping out contextually appropriate harmonization opportunities. Participants comprised national and international biobankers, clinicians, laboratory directors, bioethicists, and researchers. Key themes were identified through synthesis of participant inputs. A failure to recognize biospecimen banking as a core health research infrastructure across institutions and stakeholders was identified as a major barrier to biorepository science development in the country. This misapprehension deprioritizes national or institutional infrastructure planning, with reliance on short-term research funding for sustaining any biospecimen banking activity. The state of affairs has created siloed biospecimen collections with variable quality and limited sustainability. Regulatory oversight has followed suit with significant gaps in ethics, quality standards, and data governance. Stakeholders proposed a network-based, practitioner-led governance model as a pragmatic first step toward harmonization, enabling shared standards and capacity building in the absence of immediate legislative reform. Given the strong overlap between the science and practitioners of molecular pathology and biospecimen banking in Pakistan, incubating the field within the Association of Molecular Pathology of Pakistan was considered a practical interim solution. With very few dedicated biobanking professionals in the country, a standalone, dedicated biobanking organization is currently non-viable, but the network may transition to one as the discipline matures. The proposed model is grounded in the Pakistani context, though elements of this approach-particularly practitioner-led coordination and stepwise standardization-may be relevant to other LMIC settings with similar resource and governance constraints.
INTRODUCTION:Long-term preservation of biological material for biomedical or animal conservation purposes currently relies on ultracold temperatures. However, almost complete dehydration of cells in the presence of trehalose could allow cost-effective and flexible storage at ambient temperatures. OBJECTIVE:The study aimed to characterize optimal trehalose-loading methods before passive air-drying and explore different storage options using domestic cat primary fibroblasts as a model. METHODS:In Experiment 1, cells were loaded with trehalose using optimized electroporation (EP), thermal shock, or cold-responsive nanoparticles (CRNPs) containing trehalose. An evaluation of the cell viability after freezing and thawing was first conducted to compare trehalose protections conveyed by the different loading methods. In Experiment 2a, cells loaded with trehalose were passively air-dried for 2 days (once dry state was confirmed) and then assessed for viability, DNA integrity, or proliferative ability. In Experiment 2b, dried cells were stored for up to 4 weeks either at ambient temperatures, ambient temperatures under vacuum, 4°C, or -20°C while being assessed for viability every 24 hours for the first 5 days of storage and at the 2- or 4-week timepoint. RESULTS:Loading cells with trehalose using EP and thermal shock methods achieved cryoprotective levels, but CRNPs did not enable them to reach that. After reaching the dry state, cells from all trehalose-loading methods and untreated controls had viable cells. Additionally, trehalose-loaded cells from all methods had no increase in DNA damage after drying when compared with fresh controls. Following each method, a small number of cells retained proliferative ability. Storage at 4°C helped prolong viability through the initial 5 days compared with other storage conditions; however, no cells were viable after 2 weeks. CONCLUSION:Cells maintain viability, DNA integrity, and proliferative ability after reaching the dry state; however, storage conditions beyond 5 days must be improved.
BACKGROUND:Biobanks have evolved from simple repositories into complex research infrastructures that require robust governance frameworks to ensure traceability, data integrity, and operational reliability. ISO 20387 provides internationally harmonized accreditation standards for biobanking; however, empirical evidence evaluating its organizational impact remains limited. METHODS:We conducted a longitudinal observational study of a hospital-based biobank between 2019 and 2025. ISO 20387 accreditation obtained in 2022 was considered the structural intervention point in the analysis. Governance and quality indicators were evaluated across the sample lifecycle, including labeling error rate, metadata completeness, traceability completeness, nonconformity rate per 100 aliquots, on-time processing, and Corrective Action and Preventive Action (CAPA) closure time. RESULTS:Operational activity expanded substantially during the study period, with annual aliquot production increasing from 2142 aliquots in 2019 to a peak of 7668 in 2022. Governance indicators improved progressively during the pre-accreditation phase and stabilized following accreditation. Metadata completeness increased from 74% to 86% before accreditation to 99%-100% afterward, while traceability completeness rose from 71%-73% to 98%-99%. Nonconformity rates declined from 2.03-2.43 per 100 aliquots before accreditation to 0.75-0.90 afterward. On-time processing improved from 87%-93% to 98.5%-99%, and median CAPA closure time decreased from 53-63 to 39-42 days. CONCLUSIONS:ISO 20387 accreditation was associated with structural stabilization of governance processes in a hospital-based biobank undergoing substantial operational expansion. These findings suggest that accreditation can act as a catalyst for governance maturation, helping biobanking infrastructures maintain operational control and quality performance while scaling activity.
BACKGROUND:To investigate the effects of storage time and temperature on the quality of fecal samples and to provide a reference for clinical laboratories and biobanks in formulating sample storage operation guidelines. METHODS:Fresh fecal samples were collected from healthy volunteers and immediately aliquoted into aliquots. Different temperature and time gradients were established to simulate common pre-analytical storage processes in clinical practice, with samples snap-frozen in liquid nitrogen immediately after collection as the control group. 16S rRNA gene sequencing and untargeted lipid metabolomics were employed to determine changes in microbial diversity, species abundance, and metabolite concentrations under different storage conditions, and sample quality was evaluated based on these indicators. RESULTS:Storage at 4°C significantly minimized fluctuations in α-diversity indices, with the most pronounced protective effect observed within 2-4 hours; beyond 4 hours, changes in microbial community structure intensified. β-diversity analysis revealed that 4°C storage delayed the increase in microbial dissimilarity between samples and the liquid nitrogen-frozen control group, among which samples stored for 2-4 hours exhibited the highest similarity to the control. At the phylum level, the abundance of Firmicutes increased significantly after 6 hours of storage at room temperature, while 4°C storage effectively delayed this change. Metabolomic analysis identified more metabolomic differences (including bile acids and amino acids) in samples stored at room temperature, whereas only minor changes in fatty acid metabolites were observed at 4°C. 3β-hydroxy-5-cholenic acid exhibited a continuous upward trend with prolonged storage at both temperatures, suggesting its potential as a biomarker for evaluating sample storage quality. CONCLUSION:Short-term storage at 4°C (≤4 hours) can effectively delay the quality degradation of fecal microbial communities and metabolites, making it the optimal transitional storage strategy when immediate liquid nitrogen freezing is not feasible in clinical practice. These findings provide critical experimental data for the establishment of standardized fecal sample storage protocols.
BACKGROUND:The Kids Eye Biobank collects biospecimens, images, and clinical data from pediatric ophthalmology patients for future research use. A broad informed consent model is used; participants have opt-in choices, which include the creation of cell lines/organoids, use of whole genome sequencing (WGS) data, sharing resources with industry, and receiving future communications. This study aimed to characterize participant consent preferences and investigate associations between participant characteristics and their consent preferences. METHODS:Participant demographics, diagnosis, and consent preferences were abstracted from the Kids Eye Biobank's records. Variables were analyzed using descriptive statistics and chi-square test. Patient engagement methods were used throughout the study. RESULTS:Between January 2020 and April 2025, 391 patients were approached and 342 (87%) enrolled. Of the enrolled participants, 265 (78%) had a substitute decision-maker, and 205 (60%) were diagnosed with a malignant neoplasm affecting the eye. Demographic data were available for 273 (80%) participants; 50% were boys/men, 48% identified as belonging to a visible racial minority, and 67% reported a religious affiliation. Participants with malignant neoplasms opted in more to the use of WGS data and to the sharing of WGS data and cell lines/organoids with industry (p < 0.05). Participants belonging to a visible racial minority opted in less to the generation of cell lines/organoids (p < 0.05) and to receiving communications about future research (p = 0.002) compared with White participants. CONCLUSION:Informed consent preferences may be influenced by participant diagnosis and/or race. These findings may help the Kids Eye Biobank tailor its communication strategies and diversify its participant population.
Biobanks play a critical role in advancing biomedical research, yet they face persistent challenges related to sample findability, provenance verification, and cross-institutional collaboration. Existing systems lack standardization, suffer from data silos, and often fail to meaningfully engage donors, resulting in underutilized samples and inefficiencies. Blockchain technology, with its features of immutability, transparency, and decentralized trust, is well suited to help address these challenges. This paper explores the potential of blockchain-based decentralized biobanking, introducing key technological concepts such as distributed ledgers, smart contracts, and privacy-preserving cryptographic protocols. By enabling clear provenance trails, partially automated governance, and ethical compliance mechanisms, blockchain protocols can meaningfully address biobanking's core issues of trust, coordination, and operational complexity. We examine practical applications in improving sample visibility and governance and ensuring donor-centric ethical practices. While implementation challenges such as privacy regulations, scalability, and organizational adaptation remain, the paper argues that blockchain technology provides a robust technical framework for enhancing biobank functionality and fostering collaboration. As the field evolves, blockchain-enabled biobanking networks hold significant potential to accelerate biomedical research.
INTRODUCTION:Sperm cryopreservation induces intracellular oxidative stress. Sesamol (SE), as an antioxidant, mitigates cryodamage caused by spermatozoa cryopreservation. This study was designed to evaluate the protective effects of SE-loaded nanostructured lipid carriers (SE-loaded NLCs) on frozen sperm quality. METHODS:SE-loaded NLCs were synthesized using hot homogenization, followed by characterization for particle size, morphology, and encapsulation efficiency (EE). Normozoospermic semen samples were collected from 20 men and prepared according to the WHO 2021 guidelines. The samples were allocated into experimental groups of fresh, frozen-thawed control, SE-treated, blank NLCs, and SE-loaded NLCs. Following semen cryopreservation and thawing protocols, sperm quality was evaluated regarding motility, vitality, morphology, membrane and acrosome integrity, mitochondrial membrane potential (MMP), and DNA fragmentation using established staining and microscopy techniques. Statistical analysis was also performed at the significance level of 0.05. RESULTS:NLCs were successfully prepared with a mean particle size of 154 nm, zeta potential of -17.6 mV, and EE of 72 ± 2.64%. Fluorescence microscopy confirmed the efficient uptake of NLCs by spermatozoa within 60 minutes. The optimal SE dose was determined as 50 µM for cryoprotection. Sperm treated with SE-loaded NLCs showed improvements (p < 0.05) in motility, viability, membrane and acrosome integrity, MMP, and reduced (p < 0.05) DNA fragmentation than other groups. CONCLUSION:Application of SE-loaded NLCs showed accelerated protective effects compared with free SE, highlighting the associated potential in the enhancement of sperm quality during cryopreservation. SE-loaded NLCs effectively enhanced sperm quality indices during cryopreservation. Thus, SE-loaded NLCs represent a promising strategy to mitigate cryoinjury and improve fertility preservation outcomes.
Artificial intelligence (AI) is a powerful technology that can accelerate discovery at an unprecedented speed across all sectors of life. While some sectors already operate under established guidelines and regulations, others remain largely unchecked. With great power, however, comes great responsibility. This article, therefore, calls for a closer look at the use of AI in biobanking, a field that relies heavily on trust. AI, in return, is influenced by a plethora of interests that shape national strategies on its deployment. In relation to biobanking, political decisions play a key role in how health data are used. Ultimately, this article calls for AI governance here in the field of biobanking that employs the technology for the common good by calling for a commitment to shared responsibility by revisiting the bioethical principles of beneficence, nonmaleficence, and justice in the era of the employment of a transformative technology and its uncertain societal impacts.
