Biopreservation and Biobanking最新文献

Risk assessment represents one of the requirements for all activities involving human tissues within the premises. Although a variety of procedures are available to prepare risk assessments in general, there are no published examples of risks associated with the use of human samples in research. To cover this gap and to give an overview of the evaluation performed in our institution, we summarized the potential risks for the use of human samples in research identified in the projects under the remit of the UCL/UCLH Biobank. The procedures of acquisition, transportation, storage, use, and disposal of human samples, and security of the premises were analyzed. From our experience, there are governance-related risks associated with the process of consenting the patients, with the donor confidentiality, with mislabeling of samples and with the ethical approval associated with the project, and they generally do not compromise the integrity of the samples. On the other hand, samples' integrity is more at risk during collection, storage, transport, and use of the sample. Adequate training and having appropriate standard procedures in place and available for all staff seem to be the most effective control measures to prevent any issue. In addition, appropriate equipment maintenance, contingency plans, and strict regulation and monitoring of the facility security should always be in place. In summary, an appropriate evaluation of the risks associated with the use of human samples in research is one of the requirements for the use of human samples in research and it is fundamental for the protection of staff, students, the institution itself, and the patients. Supporting biobanking, implementing a culture of biosafety in the life sciences, and raising awareness in the scientific and regulatory communities are key ways to anticipate future problems associated with biological and governance risks.

Introduction: It is important for many research stakeholders to know how many biobanks exist. There are several potential data sources that might be expected to provide biobank numbers, such as institutions, research funders, and literature databases (e.g., PubMed), but in practice this information is rarely available and is hard to find. However, the maturation of several online health research biobank locators (also known as directories and catalogs) that relate to 12 countries and/or states has now provided some initial data to address the question of how many health research biobanks exist in relation to population size. Methods: We have analyzed four biobank locators: the Biobanking and Biomolecular Resources Research Infrastructure-European Research Infrastructure Consortium directory, the Canadian Tissue Repository Network locator, the Australian New South Wales Australia Health Pathology locator, and the UK Clinical Research Collaboration Tissue Directory. Results: We conclude that across these locators, and in those regions with potential for high research capacity as indicated by comparable gross domestic products, there are 11-30 health research biobanks/million population (2 large biobanks with >1000 samples and a further 9-28 are medium-small biobanks). Conclusion: Many locators were established primarily to increase utilization of biobanks. However, locators may be more useful in tracking the numbers of biobanks and in assisting funders and institutions to monitor research strategy and prevent unnecessary duplication of biobank resources.

Statement of the Problem: Several standards and guidelines for biobanks or biorepositories have been published by various parties (e.g., the International Society for Biological and Environmental Repositore [ISBER] and the International Organization for Standardization [ISO]). These documents are invaluable for improving the routine practices of the biobanks but the implementation has proven to be challenging for those biobanks from the non-English regions because these resources are mostly written in English. Proposed Solution: The World Health Organization (WHO) has recently published the International Classification of Diseases 11th Revision (ICD-11) along with a translation tool (lexique) for potential users. This has inspired us to make a similar contribution in the biobanking field. All the regional ambassadors (RAs) and director-at-large (DAL) in the Indo-Pacific Rim (IPR) region worked together to produce a similar lexique for potential users of ISBER's Best Practices (BPs) 4th edition. A lexique with languages of Hindi, Indonesian, Vietnamese, and Japanese has been prepared. Conclusions: This lexique is a comparison table between various languages and is expandable to other languages. In addition, this lexique will be a good tool for understanding the ISBER BPs 4th edition.

Aims: The purpose of biobanking is to provide biospecimens and associated data to researchers, yet the perspectives of biobank research users have been under-investigated. This study aimed to ascertain biobank research users' needs and opinions about biobanking services. Methods: An online survey was developed, which requested information about researcher demographics, localities of biobanks accessed, methods of sourcing biospecimens, and opinions on topics including but not limited to, application processes, data availability, access fees, and return of research results. There were 27 multiple choice/check box questions, 4 questions with a 10-point Likert scale, and 8 questions with provision for further comment. A web link for the survey was distributed to researchers in late 2019/early 2020 in four Australian states: New South Wales, Victoria, Western Australia, and South Australia. Results: Respondents were generally satisfied with biobank application processes and the fit for purpose of received biospecimens/data. Nonetheless, most researchers (n = 61/99, 62%) reported creating their own collections owing to gaps in sample availability and a perceived increase in efficiency. Most accessed biobanks (n = 58/74, 78%) were in close proximity (local or intrastate) to the researcher. Most researchers had limited the scope of their research owing to difficulty of obtaining biospecimens (n = 55/86, 64%) and/or data (n = 52/85, 60%), with the top three responses for additional types of data required being "more long term follow up data," "more clinical data," and "more linked government data." The top influence to use a particular biobank was cost, and the most frequently suggested improvement was reduced direct "cost of obtaining biospecimens." Conclusion: Biobanks that do not meet the needs of their end-users are unlikely to be optimally utilized or sustainable. This survey provides valuable insights to guide biobanks and other stakeholders, such as developing marketing and client engagement plans to encourage local research users and discouraging the creation of unnecessary new collections.

The safety of banked human adipose-derived stem cells (hADSCs) purified by 155 mM ammonium chloride (NH₄Cl)-based erythrocyte lysis has not been evaluated. This study was conducted to determine the impact of NH₄Cl-based erythrocyte lysis on the biological characteristics of cryopreserved hADSCs. Stromal vascular fractions (SVFs) were obtained from lipoaspirates and purified with NH₄Cl-based erythrocyte lysis (lysis group) or without (nonlysis group). The hADSCs were freshly isolated (fresh group) from SVFs and/or cryopreserved for 2 weeks (cryo group). The morphologies, immunophenotypes, viability, apoptosis, and growth kinetics of each group were compared. The cell cycle and differentiation capacity assays were performed in both cryopreserved groups. All groups showed similar cell morphology, immunological phenotypes, and viability. However, the main effect of lysis and its interaction with cryopreservation were observed when early apoptosis was regarded as a dependent variable in two-way repeated-measures analysis of variance. After cryopreservation, significant growth retardation and S-phase fraction reduction were observed in lytic hADSCs compared with those in nonlytic hADSCs. No significant differences in the adipogenic and osteogenic differentiation capacities were found between the two groups. Although NH₄Cl-based erythrocyte lysis did not affect the cell morphology, immunological phenotypes, viability, and adipogenic and osteogenic differentiation capacities of cryopreserved hADSCs, exposure to NH₄Cl-based erythrocyte lysis or its synergistic action with cryopreservation may induce apoptosis and inhibit the proliferation and mitosis of cryopreserved hADSCs. These results indicate that NH₄Cl-based erythrocyte lysis is not suitable for high-quality banked collection of hADSCs for future clinical applications. Further development of safe, convenient, and cost-effective purification methods of hADSCs is warranted.

Biomedical data bear the potential to facilitate personalized diagnosis and precision treatment. In the era of Big Data, high-quality annotation of human specimens has become the primary mission of biobankers, especially for tumor biobanks with large amounts of "omics" and clinical data. However, the lack of agreed-upon standardization and the gap among heterogeneous databases make information application and communication a major challenge. International efforts are underway to develop national projects on informatics management. The aim of this review is to provide references in specimen annotation to regulate and take full advantage of biological and biomedical information. First, critical data categories that are vital for specimen applications, including sample attributes, clinical data, preanalytical variations, and analytical records, are systematically listed for subsequent data mining. Second, current standards and guidelines related to biospecimen information are reviewed, and proper standards for tumor biobanks are recommended. In particular, commonly-used approaches and functionalities of data management are summarized and discussed. This review highlights the importance of informatics management of tumor specimens, defines critical data types, recommends data standards, and presents the methodologies of data harmonization for biobankers to reach high quality annotation of biospecimens.

Background: During sampling and processing, blood samples can be affected by hemolysis. Information is lacking regarding hemolysis for biobank samples. There is a need for a method that can easily measure hemoglobin as an indicator of hemolysis in stored samples before they are included in research projects. In this study we present a simple method for estimating hemolysis and investigate the effect of centrifugation speeds and temperatures on sample turbidity that commonly interferes with measurements. Methods: Using a variation of the Beer-Lambert law, we quantified the hemoglobin concentration in 75 long-term stored samples at a wavelength of 414 nm with a NanoDrop™ 8000 spectrophotometer. Owing to interference from turbidity, the samples underwent different treatments post-thawing: centrifugation at 10,000 and 20,000 g at two different temperatures (4°C and 19°C) for 15 minutes. In addition, freshly collected serum samples (n = 20) underwent a single freeze-thaw cycle, with hemoglobin measured prefreeze, post-thaw, and postcentrifugation. Kruskal-Wallis rank sum test groups and pairwise Wilcoxon rank test were used for statistical analysis. Results: A strong effect of centrifugation on the turbidity was shown for the long-term stored samples, however, this effect was independent of the temperature or centrifugation speeds. Centrifugation at 20,000 g for 15 minutes at 19°C reduced the turbidity up to 50%. A single freeze-thaw cycle in the fresh samples increased the optical density at 414 nm slightly, indicating a false increase of hemoglobin concentration. The following centrifugation reduced the concentration to less than the initial sample measurements, suggesting the presence of interference immediately after sampling. Conclusion: We describe here a simple and cost-effective NanoDrop-based method for measuring hemolysis levels intended for use in biobank facilities. We found that centrifugation, but not temperature, is a crucial step to reduce interference from turbidity.

Cryopreservation of male germline stem cells (GSCs) is an essential technique for their long-term preservation and utilization in various fields. However, the specific apoptosis pathways involved in cryoinjury during freezing remain unclear. Therefore, our study sought to identify the pathways involved in cryoinjury-induced apoptosis and thereby to improve freezing efficiency during GSC cryopreservation through the creation of a specific molecular-based cryoprotectant. The activities of caspase-8, caspase-9, caspase-3, and caspase-7 were assessed by Western blot analyses to determine the role of specific apoptosis pathways in GSC cryoinjury. Specifically, the role of a specific caspase was identified by recovery rate, relative proliferation rate, Annexin V/propidium iodide co-staining, and caspase activity using its inhibitor and activator. Moreover, the safety of the cryoprotectant was assessed by immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR). Furthermore, the efficacy of the molecular-based cryoprotectant was assessed using frozen cells in the presence of dimethyl sulfoxide (DMSO) (control), trehalose, a caspase-8 inhibitor Z-IETD-FMK [ZIF], or a mixture of the aforementioned compounds, after which the changes in Src signaling were measured. Our results demonstrated that caspase-8 plays a major role in cryoinjury-induced apoptosis and therefore its inhibition improves freezing efficiency. Specifically, a significantly higher relative proliferation rate was observed in the Z-IETD-FMK 0.01 μM-treated cells than in the DMSO control (100% ± 6.2% vs. 189.8% ± 9.5%), with decreases in both early apoptosis (16.6% ± 2.2% vs. 7.5% ± 1.0%) and caspase-8 activity (1.0-fold vs. 0.4-fold). The relative proliferation rate was significantly higher in the cryoprotectant mixture (246.0% ± 12.2%) than other individual treatment groups (trehalose 200 mM, 189.8% ± 9.5%; Z-IETD-FMK 0.01 μM, 189.7% ± 2.2%) with no significant differences in Src signaling. Therefore, our findings provide novel insights into the development of freezing protocols to enhance GSC freezing efficiency, thereby facilitating the wider adoption of GSCs in the livestock industry and/or clinical trials.