Biopreservation and Biobanking最新文献

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.

Cryopreservation of buck semen is essential in animal breeding but often damages sperm viability and integrity. The Honamli breed, a hardy Turkish goat, can benefit from improved freezing techniques using antioxidants such as Trolox (T). This study explores the effects of varying T concentrations on Honamli buck semen, assessing parameters such as motility, viability, and membrane integrity to enhance post-thaw quality. Findings support T's potential to improve semen extender formulations for preserving Honamli genetics.This study aims to freeze Honamli buck semen with T and to evaluate in vitro spermatological parameters. Three Honamli bucks, aged 2-3 years, were used in the study. Semen was collected from the bucks and mixed after removing seminal plasma. The mixed semen was diluted with a tris egg yolk extender containing three different concentrations of T (0.25 mM, 0.5 mM, and 1 mM) and control (0 mM). The diluted semen was equilibrated for 2 hours at +4 degrees and subjected to cryopreservation in liquid nitrogen vapor (-120°C for 12 minutes) and frozen. After thawing (37°C water bath for 30 seconds), the groups were evaluated at flow cytometric analysis for viability (SYBR/propidium iodide [PI]), plasma membrane acrosome integrity (FITC-PNA/PI), and mitochondrial membrane potential (JC-1), plasma membrane integrity (hypo-osmotic swelling test), microscopic evaluations for motility and morphological integrity (abnormal spermatozoa rate). The 0.5 T and 0.25 T groups showed significant improvements in motility compared with the control group (p < 0.05). The control group had the lowest plasma membrane integrity (p < 0.05). The highest morphological integrity was observed in the T groups compared with the control group (p < 0.05). In conclusion, supplementing T in buck semen extenders benefits spermatological parameters; particularly, 0.25 and 0.5 mM T could be used in Tris semen extenders during the cryopreservation process.

Introduction: Human mesenchymal stromal cells (MSCs) are attractive for both medical practice and biomedical research. Nonfreezing short-term storage may provide safe and simple transportation and promote the practical use of MSCs. Objectives: We aimed to determine the duration of efficient storage at ambient temperature (22°C) of human dermal MSCs in different three-dimensional organization and to investigate the role of cell metabolic mode in the resistance to the ambient storage damaging factors. Methods: MSCs in monolayer, suspension, and encapsulated in alginate microspheres (AMS) were stored in sealed containers at 22°С in culture medium. Viability (fluorescein diacetate /ethidium bromide) and metabolic activity (Alamar Blue assay) were assessed at 0, 3, 7, 10, and 14 days of the storage. Mitochondrial membrane potential (JC-1 test), cell cycle analysis, reactive oxygen species level, and resistance to hydrogen peroxide were analyzed under culture conditions. Results: Alginate encapsulation was shown to maintain viability (about 85%), metabolic activity, and adhesion ability during storage for 7 days. The storage of MSCs in both monolayer and suspension was less efficient. Culture of MSCs in AMS decreased basal metabolic activity, mitochondrial activity, and led to reversible cell cycle arrest compared to standard two-dimensional culture. MSCs in AMS have a lower basal level of reactive oxygen species and higher resistance to hydrogen peroxide compared with those in monolayer culture. Conclusion: Revealed shift into quiescent metabolic mode is essential for alginate-encapsulated MSCs resistance to storage at ambient temperature.