Biopreservation and Biobanking最新文献

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.

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.

Aim: Ethylene glycol (EG) has been employed as a cryoprotectant for many years in mammalian semen cryopreservation but not assessed for birds except for its recently illustrated beneficial effects on commercial chicken lines. The Indian red jungle fowl is facing trouble in its native range due to human encroachment. Therefore, the present study was designed to elucidate the cryoprotective effect of different EG concentrations (5%, 10%, 15%, and 20%) on frozen Indian red jungle fowl semen. Materials and Methods: Semen was collected from 20 cocks, and qualifying ejaculates (>70% motility) were pooled and diluted (15) with red fowl extender. EG was added to the four samples and 20% glycerol in control at 4°C. Samples were equilibrated and cryopreserved in LN₂. Semen quality and biochemical activity were assessed at various stages of cryopreservation. Results: Sperm motility, viability, plasma membrane and acrosomal integrity, chromatin integrity, and mitochondrial activity were recorded highest (p < 0.05) with 20% EG at the post-equilibration and post-thaw stages. Lipid peroxidation was recorded lowest (p < 0.05) with 20% EG compared with other concentrations and control at the post-equilibration and post-thaw stages. Conclusions: It is concluded that 20% EG exhibits cryoprotective properties in terms of regulating morphological and biochemical traits of frozen Indian red jungle fowl sperm.

Biobanking is crucial for advancing medical research and personalized medicine, offering high-quality biospecimens for studies on biomarkers, drug development, and diagnostics. Despite its global potential, challenges such as fragmented governance and varying standards hinder biorepository collaboration, particularly in South Africa (SA). A unified national biobank network could enhance research and healthcare by improving biospecimen access, ethical governance, and collaboration. Global biobank networks offer models for standardization, data sharing, and international cooperation. SA can benefit from these models by creating a centralized biobank platform, promoting capacity building, and fostering regional and global partnerships. To address the challenges SA faces regarding biobanking, the Medical Biobanks Cluster established a network named Medical Biorepositories of SA (MBirSA), which seeks to build a cohesive network of medical biorepositories in SA. Through this network, it plans to foster an inclusive culture of biospecimen and data protocol harmonization, while encouraging adherence to legal, ethical, and quality best practices and standards. The network aims to bring stakeholders together, increasing visibility and transparency, and encouraging sector-wide collaboration. MBirSA also aims to offer training to build capacity in global best practices, aid in the development of dependable biorepository infrastructure, and promote research partnerships to enhance healthcare advancements.

Introduction: Rare disease research in South Africa (SA) faces significant challenges, including limited prioritization and awareness, which hinder advancements in patient care and scientific discovery. This article explores the recruitment strategies employed by the Centre for Human Metabolomics (CHM) Biobank, a nonhospital-based academic rare disease biobank, to address these challenges. Methods: We explain the consent process and documents as well as the three recruitment models employed, namely (1) Recruitment via referring clinician, (2) implementation of monthly diagnostic follow-up sessions, and (3) recruitment of patients for specific projects through clinic-based recruitment drives. Discussion: We discuss the benefits as well as the challenges of each model. Challenges included clinician and patient time constraints, distrust from current consent practices, and limited public awareness. We elaborate on future strategies to address these gaps such as simplifying consent, expanding recruitment sites, collaborating with clinical, academic and public institutions, and raising public awareness of the role of the CHM Biobank. Conclusions: From the models employed over the past 5 years, it is evident that recruitment is most effective when patients perceive a direct benefit, such as involvement in active projects. These strategies outlined in the discussion are crucial for ensuring the CHM Biobank's sustainability, diversity, and its impact on scientific research and patient outcomes in SA.

Cryopreservation of boar semen is essential for maintaining genetic diversity and improving reproductive efficiency. However, optimizing semen extenders and packaging methods is crucial to enhancing sperm quality and cryotolerance. Equex-STM is commonly used as a surfactant to enhance cryoprotective properties by stabilizing sperm membranes during freezing and thawing. A total of 24 ejaculates (n = 24), six from each of four Hampshire crossbred boars, aged between 18 and 24 months were used in the present study. Semen was collected twice weekly by gloved hand technique. The sperm-rich fraction showing more than 70% initial motility was considered for further processing and freezing. The fresh semen was diluted in Beltsville thawing solution (BTS) at a 1:1 ratio and further processed with the addition of Fraction I and Fraction II extenders supplemented with and without 1.5% Equex-STM paste {Control (C)-no Equex; Treatment (T-1.5% Equex-STM supplementation}. The extended semen was packaged in 0.25 and 0.5 mL straws to check the effect of straw size (T₁-0.25 mL straw and T₂-0.50 mL straw). Straws were cryopreserved in liquid nitrogen (LN₂) for post-thaw sperm quality evaluation. Results showed that the sperm motility, viability, plasma membrane integrity (PMI), DNA integrity, and mitochondrial membrane potential (MMP) were significantly (p < 0.001) higher in the T group in comparison with control (C). Sperm motility, PMI, and DNA integrity were highly significant (p < 0.001) and sperm viability and MMP were significant (p < 0.05) in group T₂ in comparison with T₁.