Biopreservation and Biobanking最新文献

Background: Biobanks often lack standard mechanisms to keep donors connected to their biospecimens, reflecting a broken feedback loop that compromises trust, engagement, and scientific progress. Decentralized biobanking empowers patients to track their donations throughout the research journey, supporting personalized feedback and research collaboration via a privacy-preserving blockchain network. This case study explores operational feasibility of implementing decentralized biobanking for a large breast cancer biobank at a US academic medical center. Methods: A mixed-methods case study of the groundwork, implementation, and stakeholder feedback for a real-world decentralized biobanking app pilot was conducted. Biobank members were recruited from February to April 2023. Operational feasibility was assessed via analysis of institutional stakeholder perspectives, pilot engagement, and de-bi app activity. Findings: Physicians and other biobank stakeholders surfaced challenges surrounding managing expectations, balancing empowerment with clinical and research workflows, and navigating power dynamics between patients, physicians, scientists, and leadership. A total of 1080 participants enrolled over 10 weeks, including nearly 10% of the biobank with about 4000 biospecimens. During the pilot, biobank enrollment increased 65% versus the prior year, and there were no biobank withdrawals during or within 1 year following the pilot (p < 0.001 for both). The app, which facilitated biospecimen tracking and research engagement, was downloaded by 405 users. A total of 140 users tested the blockchain component, with 89% successfully claiming a nonfungible token representing their unique, immutable connection and access to donated biospecimens. Feedback was solicited to inform potential process improvements and risk assessments related to public relations, systems infrastructure, and ethical governance, illuminating next steps. Conclusions: We established operational feasibility for the first step toward decentralized biobanking, informed by requirements to manage expectations, workflows, and power dynamics. Our technical solution demonstrated robust participant engagement and compatibility with established biobanks, suggesting potential to build trust and align incentives and identifying next steps for communications, sustainability, and governance.

The Canadian Partnership for Tomorrow's Health (CanPath) reflects upon its original decisions around sample aliquoting strategies for its specimen inventory based on what is now commonly released to researchers. We propose an updated aliquoting strategy for new collections that balances upfront resources with volumes sought for downstream analysis. This updated aliquoting strategy will help inform teams establishing new biobanks or managing existing biobanks that are considering new collections.

Background: To systematically evaluate the long-term stability of cryopreserved RNA, we extended RNA quality monitoring in our renal biobank from 7 to 11 years for samples stored in PAXgene® Blood RNA tubes at -80°C. Materials and Methods: We assessed the suitability of archived PAXgene® RNA tubes for RNA sequencing by performing quality control on 217 chronic kidney disease samples, stratified by storage duration 7 (n = 62), 9 (n = 98), and 11 (n = 57) years. RNA was extracted from 2.5 mL whole blood using the PAXgene® Blood RNA Kit, with quality assessed based on concentration (Qubit™ Fluorometer, yield), purity (NanoDrop™ 2000 spectrophotometer, A_260/A280 and A_260/230 ratios), and integrity (Agilent 2100 Bioanalyzer, RNA integrity number, RIN). Sequencing eligibility required ≥500 ng total RNA and RIN ≥6.0. Results: Median RNA yields were comparable across storage durations (7-year 7.00 µg, 9-year: 7.11 µg, and 11-year: 6.79 µg, p = 0.870). The median A_260/280 ratios were 2.03 (7-year), 2.08 (9-year), and 2.07 (11-year) (p < 0.001, all ≥1.8), while median A_260/230 ratios were 1.72, 1.77, and 1.87, respectively (p = 0.550). RNA integrity, as measured by RIN, showed median values of 8.90 (7-year), 9.00 (9-year), and 8.80 (11-year). While no significant differences were observed between the 7- and 9-year (p = 0.537) or 7- and 11-year groups (p = 0.052), the 9-year group had slightly higher RIN values than the 11-year group (p < 0.05). Sequencing suitability remained consistently high (7-year: 97%, 9-year: 98%, and 11-year: 98%, p = 0.750), with 98% (212/217) of samples meeting the standards. Even under stricter RIN thresholds, pass rates remained robust (RIN ≥ 7.0: 94%, RIN ≥ 8.0: 88%). Conclusion: PAXgene® Blood RNA tubes stored at -80°C for up to 11 years provide high-quality RNA suitable for total RNA sequencing.

Introduction: Optimal cryopreservation of testicular tissue is essential for species research and conservation, enabling long-term storage of genetic resources through vitrification or slow freezing. Comparing responses from different taxonomic groups to these techniques is crucial for refining protocols and improving cryopreservation outcomes. Objectives: This study evaluated the effects of cryopreservation on cell viability, morphology, mitochondrial activity, and proliferative potential to optimize testicular tissue preservation strategies for wildlife conservation. Methods: We assessed testicular tissue from Felidae and Cervidae, including domestic cats (Felis catus) and Neotropical deer species, white-tailed deer (Odocoileus virginianus), Brazilian red brocket deer (Mazama nana), and gray brocket deer (Subulo gouazoubira). Experimental groups included control (no cryopreservation), slow freezing using Mr. Frosty or progressive temperature decrease, and conventional vitrification. Results: All methods preserved live cells and normal tissue morphology; however, compared with fresh tissue, cryopreservation significantly reduced tissue viability, mitochondrial membrane potential, and the proportion of intact seminiferous tubules. Species-specific differences emerged, with vitrification being most effective for domestic cats, while slow freezing yielded better results for Neotropical deer. Despite lower viability scores, vitrification could still be an acceptable option for cervids due to its rapid processing and minimal equipment requirements. In addition, post-cryopreservation tissue culture increased cell abnormalities, highlighting the need to optimize culture conditions for different species. Conclusion: This comparative study advances reproductive tissue preservation techniques and emphasizes the importance of tailored cryopreservation as well as in vitro culture protocols for diverse taxonomic groups.

Plant germplasm is critical for maintaining biodiversity and mitigating genetic erosion. While cryopreservation has emerged as an increasingly vital approach for conserving plant genetic resources, its routine application remains limited due to species- and explant-specific preservation requirements. In many instances, additional experimentation is necessary to develop genotype-specific protocols to achieve satisfactory survival rates. In addition, the technical operation is time-consuming and labor-intensive. Moreover, the survival rate of explants post-cryopreservation from many species remains suboptimal. PVS-based vitrification cryopreservation of dormant buds represents an ideal strategy to address these challenges, leveraging the buds' inherent cold hardiness and robust regrowth capacity. This review focuses on four key determinants of successful PVS-based cryopreservation: the collection time and physiological status of dormant buds, shoot tip dissection, PVS exposure duration, and recovery culture. The insights provided herein may facilitate the rapid development of new protocols or user-friendly procedures for a broader range of plant species, enabling wider adoption by institutions and laboratories, and thereby enhancing global efforts in germplasm conservation.

Introduction: The province of Misiones, located in north-eastern Argentina, harbors one of the best-preserved remnants of the Atlantic Forest, which is a global biodiversity hotspot. However, 95% of this forest has been deforested, posing a significant threat to numerous species, particularly to mammals. To conserve biodiversity, the Misiones' Institute of Biodiversity, which is developing a biobank, and the Center for Rescue, Rehabilitation, and Recovery of Wildlife Güirá Oga (GO), dedicated to the care of injured and displaced fauna, were established. This article highlights the collaboration between these two institutions aimed at strengthening mammal conservation efforts in Misiones. Materials and Methods: A standardized protocol was established for the collection, preservation, and storage of biological samples, considering variables such as the taxonomic group, animal condition, and study type. Samples were collected under anesthesia and in accordance with animal welfare guidelines, and relevant data were recorded. Within the Biobank, samples were classified, documented, and stored at -80°C, 4°C, or room temperature, depending on their characteristics. Biobank operations comply with international regulations-including the Convention on Biological Diversity and the Nagoya Protocol-as well as provincial legislation. Access permits and Material Transfer Agreements (MTA) are applied for any external use. Results: Between 2020 and 2024, biological samples were collected from 335 individual native mammals representing 38 species, of which 37.9% were classified as threatened (vulnerable, endangered, and critically endangered) according to their conservation status. Biobank stores 985 samples 53.5% blood and derivatives, 6.7% tissues, 11% DNA/RNA, 17% ectoparasites, and 8.2% endoparasites. They also include swabs, fibroblasts, hair, and leather. The most represented orders were Carnivora, Primates, and Didelphimorphia (63.6%). Conclusions: The collaboration between GO and the Biobank integrates rescue, research, and genetic conservation, optimizing resources and strengthening capacities to address biodiversity loss, carrying out epidemiological surveillance, and promoting future ecological restoration projects.

Introduction: The effect of prolonged storage (12-19 years) on selected laboratory test results is examined in the National Health and Nutrition Examination Survey biospecimen collection to determine whether biospecimens stored long term in vapor-phase liquid nitrogen provide valid results once remeasured. Methods: Biospecimens were selected for remeasurement using systematic random sampling for five analytes: cotinine, methylmalonic acid (MMA), vitamin A, vitamin E, and hepatitis C virus RNA (HCV-RNA). Measurements from the original specimens in 1999-2000 or 2005-2006 are compared with 2018-2019 measurements from the same survey participants and specimens. For quantitative analytes, measurement accuracy is assessed using standard method comparison procedures, precision is evaluated by comparing to quality control standards, and reproducibility is estimated by treating data like an incurred sample reanalysis. Qualitative measures are analyzed using concordance measures and exact binomial tests. Results: Observed proportional differences are 3%-12% for cotinine in people who do not smoke, 11% for cotinine in people who smoke, -8% to 1% for vitamin A, 8%-9% for vitamin E, and -6% to 8% for MMA. Precision estimates are within the standards established by quality control data and generally applicable quality goals. Differences between measurements are within 20% of the average value for at least 85% of all samples. For qualitative HCV-RNA and MMA results, we observe 99% concordance between measurements. Conclusions: Multipronged analysis showed that most differences are within acceptable ranges based on standard laboratory criteria for assessing accuracy, precision, and reproducibility. Results suggest future measurements and subsequent statistical analyses of stored serum specimens should be valid.

Objectives: Personalized medicine emphasizes prevention and early diagnosis by developing genetic screening and biomarker assessment tools. Biobanks, including University of Piemonte Orientale (UPO) Biobank, support this effort by providing high-quality biological samples collected, processed, and stored using optimized standardized protocols. To determine the optimal long-term storage conditions for biospecimens used in biomedical research, we evaluated plasma and serum samples cryopreserved using two storage methods, cryovials and straws, across various analytical methodologies with differing sensitivity and robustness. Design and Methods: Plasma and serum samples cryopreserved in liquid nitrogen in vials and straw at the UPO Biobank were subjected to multiple analyses including standard biochemical laboratory analysis, targeted lipidomics, untargeted proteomics, and targeted metabolites quantification through mass spectrometry-based analytical techniques. Results: Our data demonstrate the robustness and applicability of both storage methods for standard laboratory analyses in evaluating clinically relevant markers in plasma and serum. Lipidomic analysis revealed slight disparities in lipid abundance, though these differences were mostly confined to specific lipid species, particularly fatty acids. Conversely, proteomic and metabolomic analyses uncovered variations in abundance in a significant, albeit limited, fraction of analytes between vials and straw-derived samples. Conclusions: By highlighting similarities and differences in samples stored in these conditions, this study provides significant insights into optimizing biobanking practices and understanding the factors that influence the integrity of cryopreserved biospecimens and the reliability of the data derived from them. Both straws and vials are convenient and efficient cryopreservation methods, essentially equivalent for samples dedicated to robust and relatively low-sensitive standardized analyses. However, our findings emphasize the need for caution when interpreting omics data from samples subjected to different cryopreservation methods, as subtle variations can arise even with different types of containers.