Biopreservation and Biobanking最新文献

Reactive oxygen species (ROS) during cryopreservation causes mechanical, biochemical, and structural damage to the sperm, which leads to reduced sperm motility and fertility. N-acetyl cysteine is a cysteine-derived amino acid antioxidant that functions as a scavenger of ROS and regulates mitochondrial activity. Mitochondrial uncoupling protein 2 (UCP2) plays a leading role in this process and is one of the major regulators of human spermatozoa motility and metabolism. The purpose of the study was to examine the changes in UCP2 in frozen-thawed human sperm when exposed to N-acetyl cysteine, an effective antioxidant commonly used in human semen freezing. Semen samples were collected from 20 normozoospermia men and were divided into four experimental groups: fresh, frozen control, frozen N-Acetylcysteine (NAC, 100 μM), and frozen negative control with Genipin (25 μM). Subsequently, post-thaw sperm quality parameters, as well as UCP2 relative quantity, ROS, mitochondrial membrane potential (MMP), and malondialdehyde, were assessed. Semen treated with NAC exhibited significantly higher total and progressive motility, as well as viability, when compared to the control and genipin groups (p < 0.05). Moreover, UCP2 relative quantity was significantly lower in all frozen groups compared to the fresh group (p < 0.0001). The UCP2 relative quantity was not significantly different between NAC and control groups (p ≥ 0.05). Also, there were no significant differences in MMP, ROS, and malondialdehyde levels among the frozen groups (p ≥ 0.05). It can be concluded that UCP2 undergoes a modification during cryopreservation, and it could be an explanation of the reduction in post-thaw motility of sperm. Additionally, NAC supplementation in freezing media enhances post-thaw sperm motility and viability.

Introduction: The development of ice crystals during the freezing process can be detrimental to the viability and fertilization capacity of frozen-thawed spermatozoa. The unique properties of antifreeze proteins allow them to inhibit the formation of ice crystals during cell cryopreservation.

Objective: Previous studies have assessed the general sperm quality parameters, reactive oxygen species (ROS) level, lipid peroxidation, and sperm apoptosis after using antifreeze protein III (AFP III) for cryopreservation. However, the data regarding changes in protein expression and their relation to sperm quality after thawing are still lacking. Therefore, this work addresses associated proteomic changes in post-thawed dog sperm.

Methods: Two experiments were conducted using high (Experiment I) and low (Experiment II) concentrations of AFP III. Semen samples from four dogs were divided into aliquots and diluted with Tris-egg yolk extender supplemented with 0 (control), 1, 5, 10, or 15 µg/mL AFP III (Experiment I) or 0, 1, 2, 3, or 4 µg/mL AFP III (Experiment II) based on a previous literature review. After being frozen in LN₂ and thawed, sperm motility parameters, viability, acrosome integrity, and apoptosis were evaluated. Furthermore, the AKAP4, ATP1B1, and HSP70 proteins, which are associated with good sperm quality and freezability, were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting to assess their expression and levels.

Results: In the high-concentration experiment, AFP III at 1 µg/mL significantly increased (p < 0.05) total and moderate-progressive motility compared with the control and other AFP III concentrations. Moreover, the 1 µg/mL group had higher HSP70 and AKAP4 protein levels than the control and other concentration groups, though the differences were not significant.

Conclusion: These findings suggest that adding 1 µg/mL AFP III to the semen extender enhances the motility of post-thawed dog sperm and suggest the potential use of HSP70 and AKAP4 proteins as biomarkers for good semen quality.

Biobanks serve as the cornerstone of translational research. Evolving from traditional biobanks, living biobanks-particularly organoid living biobanks-have emerged as a critical and powerful platform, characterized by their three-dimensional biomimetic architecture, long-term self-renewal capacity, and retention of key genetic and pathological phenotypes of the parental tissue. At the pivotal juncture of a paradigm shift in biomedical research, organoids, as an important component of Novel Alternative Methods, hold broad prospects for both biomedical research and clinical applications. High-quality organoids can precisely recapitulate the structure and function of native organs, ensuring the accuracy and reproducibility of research outcomes. This establishes a robust foundation for investigating disease mechanisms, drug discovery, and precision medicine. Implementing rigorous quality control is therefore pivotal for guaranteeing research reliability and clinical applicability. The present article comprehensively examines the current landscape of tumor organoid quality control, covering critical quality control checkpoints across key technical stages of the construction process, advancements in standardization, and future development trends. By synthesizing these aspects, this work aims to empower researchers and practitioners to overcome challenges in quality control, enhance organoid fidelity, and accelerate the translation of organoid technology from fundamental research to clinical implementation.

Background: A widely representative health system cohort with longitudinal specimen collection can serve as an efficient clinical biobank resource for multiple studies. Because the full scope of a health system cohort can include both health care workers and patients, enrollment and biobanking efforts may be designed to engage these specific participant populations. Methods: For a multisite health system cohort that initially enrolled health care workers and then expanded to enroll patients, we evaluated the relative success of initiatives that specifically targeted enrollment of various health care worker and patient populations. We also compared enrollment rate success based on engagement type (active vs. passive), modality (in-person vs. virtual), and venue (clinical-based or community-based). Across each method of engagement, we compared the conversion rate from study consent to collected biospecimen. Results: For recruitment activities involving health care workers, enrollment rates varied based on active versus passive (62% vs. 0.8%) and in-person versus virtual (9.6% vs. 0.8%) engagement as well as clinical-based versus community-based (65% vs. 3.9%) venues (p < 0.001 for all). For health care workers, the overall conversion rate from consent to biospecimen collection was 87%. For recruitment activities involving patients, enrollment rates also varied based on active versus passive (53% vs. 0.8%) and in-person versus virtual (62% vs. 0.8%) engagement, as well as clinical-based versus community-based (70% vs. 41%) venues (p < 0.001 for all). For patients, the overall conversion rate from consent to biospecimen collection was 75%. Conclusions: For studies aiming to build a biorepository resource involving both health care worker and patient participants, the active rather than passive engagement methods are likely to achieve not only a higher rate of contact to consented enrollment but also a higher rate of conversion from consent to biospecimen collection. Further studies are needed to guide resource planning around biorepository building capacity for specific study designs.

Introduction: Extracellular vesicles (EVs) are lipid bilayer particles released by all cell types, carrying cargos that reflect the cellular states of their origin. Recently, EVs are increasingly recognized as valuable biomarkers and therapeutic vectors in oncology, but their clinical translation is limited by variability in isolation methods and uncertainty regarding long-term storage physical stability. Methods: We systematically compared human plasma EVs isolated by ultracentrifugation (UC) or magnetic bead (MB)-based methods under immediate analysis, stable freezing storage, and repeated freeze-thaw conditions. The morphology and protein profiling of EVs were characterized by transmission electron microscopy (TEM) and Western blotting (WB), respectively. EV concentration, particle size, and zeta potential were quantified by particle size analyzer. Results: TEM and WB analyses of human plasma EVs confirmed the efficacy of both the UC and MB isolation methods. UC-isolated EVs are of high yield but low physical stability, featuring size reduction and a shift toward more negative zeta potential values after freeze-thaw cycles. Fresh UC-EVs displayed heterogeneous size profiles, whereas freeze-thawed samples shifted to a dominant peak, consisting of small particles with increased counts. Although lower in yield, MB-isolated EVs retained their physical stability across all conditions. Conclusion: MB-based EV isolation offers physical stability for standardized diagnostic workflows, whereas UC-based EV isolation provides high yield for discovery studies but vulnerable to freeze-thaw stress. These findings provide an evidence-based framework for selecting EV isolation and storage methods to match downstream applications, guiding the standardization of EVs workflows for future precision oncology and personalized medicine. [Figure: see text].

Latin America hosts extraordinary biological diversity but remains underrepresented in global biomedical research, underscoring the need for robust biobanking infrastructures. This work provides an updated snapshot of Chilean biobanks, based on a national survey exploring their current capacities and challenges. Nine active biobanks were identified across 5 of Chile's 16 regions, the majority concentrated in Santiago. Collectively, they store over 640,000 biospecimens from nearly 49,000 participants, predominantly oncological. While standardized protocols for sample management are broadly implemented by Chilean biobanks, data management practices are not yet well-developed, as only a few centers have adopted internationally recognized standards. Governance structures vary considerably and often lack formal written documentation. Financial sustainability relies mainly on institutional support, competitive grants, and modest cost recovery. Although Chilean biobanks contribute to research and training, measuring productivity remains challenging due to underreported acknowledgments and limited post-transfer traceability. Overall, our analysis suggests a bottom-up development of Chilean biobanks in the absence of dedicated legislation or strategic governmental policies. This overview shows that Chile's biobanks hold considerable potential for strengthening translational research and health equity, particularly if further support enables expansion into underrepresented regions. By integrating these infrastructures into higher education, clinical care, and broader regional collaborations, biobanks can help leverage Chilean genetic diversity and address health disparities. With greater governmental prioritization, a cohesive regulatory framework, and collaboration as a key strength, biobanks could enhance interaction with global networks and further strengthen Latin America's overall contribution to biomedical innovation.

In June 2021, Argentina's Ministry of Science, Technology, and Innovation launched PoblAr-the Program of Genomic Reference and Biobank of the Argentinian Population. This pioneering initiative aims to generate representative human genomic data and associated metadata for Argentina, a crucial step toward advancing genomic research and public health in the country. PoblAr addresses a significant knowledge gap in a country with a rich and dynamic history of population admixture, where unique genetic and environmental diversity shape health and disease patterns. As one of Latin America's first large-scale genomic initiatives, PoblAr aligns with similar efforts in Mexico and Brazil, reinforcing its regional and global relevance. The program's comprehensive sampling protocols integrate biological and nonbiological traits, enabling a multidimensional biobank designed to identify statistical risk factors across diverse conditions. A robust ethical framework underpins PoblAr, prioritizing donor safety, data confidentiality, and equitable community benefits through rigorous informed consent and governance tailored to its scale. PoblAr has established a secure data infrastructure using local informatics tools and enforcing strict anonymization protocols through multilevel access controls. Recent studies on local samples reveal that Argentina's ancestral composition is more complex and nuanced than previously reported. The program places a strong emphasis on community engagement through an exhaustive communication strategy that fosters collaboration with donors, the broader public, and local governments. By promoting data-driven precision health initiatives across Argentina, PoblAr aims to deliver significant societal benefits and encourage inclusivity.

The Neotropical region is currently facing a critical period of biodiversity loss, with deer species (family Cervidae) being particularly affected by severe habitat degradation, genetic bottlenecks, and population fragmentation. In this context, germplasm biobanks emerge as strategic tools for conservation efforts. This article presents a comprehensive review of the Deer Research and Conservation Center germplasm bank, the largest repository of genetic material for Neotropical deer worldwide. We detail the diversity of species represented, the types and quantities of cryopreserved samples, and the operational costs associated with maintaining the biobank. Additionally, we discuss the main advantages of germplasm banking, such as preserving genetic diversity without the logistical challenges of managing large captive populations, as well as critical challenges, particularly those arising from ongoing taxonomic uncertainties that complicate species identification and sample management. Scientific applications and conservation actions already enabled by this resource are presented, alongside a discussion of future perspectives, including potential expansions of sample types and integrative genomic analyses. This review underscores the essential role of germplasm biobanks in preserving the genetic legacy of Neotropical deer and supporting long-term biodiversity conservation strategies.