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Artificial Intelligence-Based Quality Control of Cell Lines. 基于人工智能的细胞系质量控制。
IF 1.4 4区 生物学 Pub Date : 2025-08-12 DOI: 10.1177/19475535251367317
Svetlana Gramatiuk, Igor A Kryvoruchko, Yulia V Ivanova, Emily Hubbard, Maria Noebauer-Babenko, Karine Sargsyan

Introduction: This study is part of the broader Stem Line project Mito-Cell-UAB073, specifically focusing on "Stem Cell Lines-Quality Control," and aims to innovate in the field of Quality Control (QC) through a unique, artificial intelligence (AI)-powered model known as Life Cell AI UAB. This model utilizes deep learning algorithms and computer vision, allowing it to make accurate viability assessments of cell and stem cell lines based solely on static images captured through standard optical microscopes. Aim: The aim of this study was to develop and validate an AI-driven, image-based model that reliably predicts cell line viability. Methods: Our methodology involved training the Life Cell AI UAB model on single static images of cell lines using advanced computer vision and deep learning techniques. Performance evaluation was conducted on three independent blind test sets sourced from various biotechnology laboratories, allowing for assessment across diverse environments. Results: The Life Cell AI UAB model achieved a sensitivity of 82.1% in identifying viable cell lines and a specificity of 67.5% for non-viable lines across the test sets. Each blind test set exhibited a weighted accuracy above 63%, with a combined accuracy of 64.3%. Notably, predictions showed a clear distinction between correctly and incorrectly classified cells. The model outperformed traditional QC methods by improving accuracy in binary classification tasks by 21.9% (p = 0.042) and demonstrated a 42.0% enhancement over conventional Standard Operation Procedure (SOP) procedures (p = 0.026). Conclusion: The Life Cell AI UAB model represents a notable advancement in biobanking QC, offering a precise, standardized, and non-invasive method for assessing cell line viability. This model has the potential to streamline QC processes across laboratories, minimizing the need for time-lapse imaging and promoting uniformity in QC practices for both cell and stem cells.

本研究是更广泛的干细胞系项目Mito-Cell-UAB073的一部分,特别关注“干细胞系-质量控制”,旨在通过一种独特的人工智能(AI)驱动的模型,即生命细胞AI UAB,在质量控制(QC)领域进行创新。该模型利用深度学习算法和计算机视觉,使其能够仅根据通过标准光学显微镜捕获的静态图像对细胞和干细胞系进行准确的活力评估。目的:本研究的目的是开发和验证一个人工智能驱动的、基于图像的模型,该模型可以可靠地预测细胞系的生存能力。方法:我们的方法包括使用先进的计算机视觉和深度学习技术在细胞系的单个静态图像上训练Life Cell AI UAB模型。性能评估是在三个独立的盲测试集上进行的,这些盲测试集来自不同的生物技术实验室,允许在不同的环境中进行评估。结果:Life Cell AI UAB模型在识别活细胞系方面的灵敏度为82.1%,在识别非活细胞系方面的特异性为67.5%。每个盲测集的加权准确率均在63%以上,综合准确率为64.3%。值得注意的是,预测显示了正确和错误分类细胞之间的明显区别。该模型在二元分类任务中的准确率比传统QC方法提高了21.9% (p = 0.042),比传统的标准操作程序(SOP)程序提高了42.0% (p = 0.026)。结论:Life Cell AI UAB模型代表了生物银行QC的显著进步,为评估细胞系活力提供了一种精确、标准化和无创的方法。该模型具有简化实验室QC流程的潜力,最大限度地减少了对延时成像的需求,并促进了细胞和干细胞QC实践的统一性。
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引用次数: 0
The College of American Pathologists Biorepository Accreditation Program: Results from the First 10 Years. 美国病理学家学院生物库认证计划:前10年的结果。
IF 1.4 4区 生物学 Pub Date : 2025-08-12 DOI: 10.1177/19475535251365758
Nalin Leelatian, Joan H Rose, Sarah Cliff, Richard C Davis, Helena Ellis, Jing Di, Basal Kashlan, Nilsa C Ramirez, Jim Vaught, Erik Zmuda, Shannon J McCall, Rebecca C Obeng

Introduction: The College of American Pathologists (CAP) Biorepository Accreditation Program (BAP) was established in 2012 with the goal of providing standardized requirements that ensure high quality for procuring, processing, storing, distributing, and computerizing information of biospecimens for scientific investigations. CAP BAP was the first biorepository accreditation program, and, since the program started in 2012, the world's second biorepository accreditation standard was issued by the International Organization for Standardization (ISO) as ISO 20387 in 2018. CAP BAP serves as an interface between several programs and draws best practices from renowned organizations. This elective program is based on a peer-inspection model to ensure that the inspectors have proper expertise and to promote educational efforts through information sharing. On-site inspections occur every 2 years, like other CAP Accreditation Programs, with an interim self-inspection in the off year. The program compliance is assessed based on CAP Accreditation Checklists, which are regularly revised. Objective: This article reviews the accomplishments of the first 10 years of the CAP Biorepository Accreditation Program. Results: As of December 2024, 104 biorepositories are CAP BAP accredited, which increased from 53 accredited biorepositories in 2018. Accreditation of 10 additional biorepositories is underway. A total of 88 inspections were completed between January 2017 and December 2022; 16 were initial inspections and 72 were reinspections. Deficiencies, defined as insufficient or lack of evidence of compliance with a checklist item, were mainly related to equipment/instrumentation (24%), quality management (15%), safety (14%), information technology (13%), personnel (13%), specimen handling and quality control (9%), facilities (6%), and regulatory (6%) issues. The proportion of deficiencies between categories was like the first 5 years. Conclusion: The increased number of accredited biorepositories, in both academic and commercial settings, highlights the continued success of the program and its applicability to maintaining high standards for biorepositories.

简介:美国病理学家学会(CAP)生物库认证计划(BAP)成立于2012年,旨在提供标准化要求,确保科学调查生物标本的采购、处理、存储、分发和计算机化信息的高质量。CAP BAP是第一个生物储存库认证计划,自2012年该计划启动以来,国际标准化组织(ISO)于2018年发布了世界上第二个生物储存库认证标准ISO 20387。CAP BAP是多个项目之间的接口,并借鉴了知名组织的最佳实践。这个选修课是基于同行检查模式,以确保检查员有适当的专业知识,并通过信息共享促进教育努力。与其他CAP认证项目一样,每两年进行一次现场检查,并在非年度进行一次临时自检。该计划的合规性是根据定期修订的CAP认证清单进行评估的。目的:本文回顾了CAP生物库认证计划前十年的成就。结果:截至2024年12月,获得CAP - BAP认证的生物储存库从2018年的53个增加到104个。正在对另外10个生物储存库进行认证。2017年1月至2022年12月期间共完成了88次检查;16次为初次检查,72次为复检。缺陷,定义为不充分或缺乏符合清单项目的证据,主要与设备/仪器(24%)、质量管理(15%)、安全(14%)、信息技术(13%)、人员(13%)、标本处理和质量控制(9%)、设施(6%)和法规(6%)问题有关。不同类别之间的缺陷比例与前5年相似。结论:在学术和商业环境中,获得认证的生物储存库数量的增加,突出了该计划的持续成功及其在保持生物储存库高标准方面的适用性。
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引用次数: 0
Transforming Biospecimen Management: A Roadmap for Integrated Sample Traceability in the Era of Global Research. 转变生物标本管理:全球研究时代集成样本可追溯性的路线图。
IF 1.4 4区 生物学 Pub Date : 2025-08-12 DOI: 10.1177/19475535251366364
Sion Israel Sion, Trinh Nguyen-Phan, Mélissa Fortin, Anne-Marie Mes-Masson, Kaiwen Zhang

Introduction: Advancements in biomedical research depend on the quality and availability of biological samples. Despite their sophisticated storage capabilities, biobanks face significant challenges in sample management, with stored specimens often remaining unused and researchers struggling to access the required samples. Objectives: To analyze the challenges in biospecimen access and traceability, evaluate existing solutions, and propose a framework for integrated sample management in global research collaboration. Methods: A scoping review was conducted across PubMed, Scopus, and Web of Science databases, supplemented by grey literature (2004-2024). The analysis included an examination of Biobank Information Management Systems and an evaluation of sample management systems, tracking technologies, and governance frameworks. Results: The analysis revealed fragmented management systems, with at least 38 different biobanking software solutions offering limited interoperability. Proprietary systems and vendor lock-ins create significant barriers to data sharing. Sample tracking shows the evolution from manual to digital systems; however, cross-institutional tracking remains challenging. Reproducibility issues account for significant challenges in research, whereas inefficient resource utilization persists, with 67% of biobanks citing underutilization as a major concern. Conclusions: Addressing biobank sample access and traceability requires a shift from an institution-centric to an ecosystem-wide approach. Its success depends on integrating technological solutions such as Blockchain, the Internet of Things, and artificial intelligence with governance frameworks while ensuring alignment with stakeholder needs. Future developments should focus on implementing integrated traceability systems that support transparent and accountable sample management across the global research ecosystem.

生物医学研究的进步取决于生物样品的质量和可用性。尽管生物库具有复杂的存储能力,但在样本管理方面面临着重大挑战,存储的样本通常未被使用,研究人员很难获得所需的样本。目的:分析生物标本获取和可追溯性方面的挑战,评估现有解决方案,并提出全球研究合作中综合样本管理的框架。方法:对PubMed、Scopus和Web of Science数据库进行范围综述,并辅以灰色文献(2004-2024)。分析包括对生物样本库信息管理系统的检查和对样本管理系统、跟踪技术和治理框架的评估。结果:分析揭示了分散的管理系统,至少有38种不同的生物银行软件解决方案提供有限的互操作性。专有系统和供应商锁定为数据共享造成了重大障碍。样本跟踪显示了从手工系统到数字系统的演变;然而,跨机构跟踪仍然具有挑战性。可重复性问题是研究中的重大挑战,而资源利用效率低下仍然存在,67%的生物库认为利用不足是一个主要问题。结论:解决生物样本获取和可追溯性问题需要从以机构为中心的方法转向全生态系统的方法。它的成功取决于将区块链、物联网和人工智能等技术解决方案与治理框架相结合,同时确保与利益相关者的需求保持一致。未来的发展应侧重于实施集成的可追溯系统,以支持整个全球研究生态系统中透明和负责任的样品管理。
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引用次数: 0
Data Governance and Distribution of Biobank: A Case from a Chinese Cancer Hospital. 生物样本库的数据治理与分布:以某肿瘤医院为例。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2024-12-13 DOI: 10.1089/bio.2024.0081
Jingjing Shi, Yan Guo, Na He, Wenbin Xia, Hongkun Liu, Haixin Li

Objectives: To facilitate the regionalization, specialization, and digitization of biobanks, three issues regarding data collection and application must be addressed (1) integration and distribution of data governance, (2) efficiency and efficacy of data governance, and (3) sustainability of data governance. Methods: We collaborated with stakeholders to identify priorities and assess infrastructure needs through the continuous evaluation and analysis of projects. We developed data management solutions, catalogs, and data models to optimize and support data collection, distribution, and application. Furthermore, ontologies were used to facilitate data integration from multiple sources, and Minimum Information About BIobank Data Sharing (MIABIS) was defined as accessible to all patients. To enhance data integrity, we conducted retrospective and prospective follow-up studies. Results: We completed infrastructure upgrades to match technical solutions and research demands. An information management software with six primary functional divisions was developed for data governance. We optimized the database structure and changed the biospecimen accumulation model from biospecimen-based to patient-centered and service-oriented. Subsequently, we specified 85 attributes of MIABIS to describe the biobank contents. A dual-pillar approach was adopted to expand the biobank's data in collaboration with other institutions, and MIABIS served as a bridge for both vertical and horizontal networks. From 2003 to 2021, we collected a total of 156,997 patient biospecimens/data from 20 cancer types, matching 53,113 cases from follow-up surveys. In addition, we supplied more than 40,000 biospecimens/data points for above 300 scientific research projects. Conclusions: An appropriate information platform for a biobank is fundamental to data collection, distribution, and application, particularly in the context of data-intensive research. We implemented a standardized scientific data structure to fulfill the research requirements. The sustainable development of a biobank depends on a scientific, standardized, and service-oriented data governance approach, along with the efficient utilization of emerging technologies.

目标:促进生物库区域化、专业化和数字化,必须解决数据收集和应用中的三个问题:(1)数据治理的整合和分布;(2)数据治理的效率和功效;(3)数据治理的可持续性。方法:我们与利益相关者合作,通过对项目的持续评估和分析,确定优先事项并评估基础设施需求。我们开发了数据管理解决方案、目录和数据模型,以优化和支持数据收集、分发和应用。此外,本体论用于促进来自多个来源的数据集成,并将生物银行数据共享的最小信息(MIABIS)定义为所有患者都可以访问。为了提高数据的完整性,我们进行了回顾性和前瞻性随访研究。结果:我们完成了基础设施升级,以匹配技术解决方案和研究需求。针对数据治理,开发了具有六个主要功能部门的信息管理软件。优化数据库结构,将生物标本积累模式从“以生物标本为基础”转变为“以患者为中心、以服务为导向”。随后,我们指定了85个MIABIS属性来描述生物库内容。采用双支柱方法与其他机构合作扩大生物库的数据,而MIABIS则作为垂直和水平网络的桥梁。从2003年到2021年,我们共收集了来自20种癌症类型的156997例患者生物标本/数据,与随访调查的53113例病例相匹配。此外,我们还为300多个科研项目提供了4万多个生物标本/数据点。结论:一个合适的生物库信息平台是数据收集、分发和应用的基础,特别是在数据密集型研究的背景下。实施规范的科学数据结构,满足科研需求。生物库的可持续发展有赖于科学、规范、服务型的数据治理方法,以及新兴技术的高效利用。
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引用次数: 0
Challenges and Opportunities for Collaboration Between Academic Biobanks and Industry: Results of an International Survey of Academic Biobanks. 学术生物库与产业界合作的挑战与机遇:学术生物库国际调查的结果。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2024-11-06 DOI: 10.1089/bio.2023.0156
Wohaib Hasan, Daniel Simeon-Dubach, Vanessa Tumilasci, Peter Sebbel, Suenne Orth

Aim of the Survey: When it comes to collaboration between academic biobanks and the pharmaceutical/biotechnology industry, the criteria for effective collaborations are still unclear. Researchers in industry and academic biobanks can have different incentives and requirements that the other party is often not familiar with. This survey was conducted in an attempt to increase understanding of these fundamental knowledge gaps that may be obstacles to optimal collaboration between academia and industry. Key Findings from the Survey: There were 53 total respondents. Although this was a global survey, most respondents (n = 29) were from North America, likely reflecting overall investment in research in this region and possibly increased interactions between academia and industry as well. Most respondent academic biobanks collect multiple sample types with most (>90%) collecting both biofluids (including blood) and tissue. Most of the participating academic biobanks were aware that they were not (35%), or only partially (35%), using the full potential of their inventory. One option for increasing utilization rates is by collaborating with industry partners. The main issues when working with industry were perceived to be a combination of challenges including contractual (55%), consent restrictions (45%), timelines (41%), or time pressure (36%). Time taken to put agreements together was also a significant hurdle (54%), together with the industry's administrative requirements (36%). Brief Conclusions from the Survey: To take advantage of opportunities for joint collaboration, it is essential that the parties involved build trust. The first step is to understand the different requirements and needs of the other party and to establish efficient structures for joint cooperation. This survey has highlighted key areas to be addressed as the next steps for strengthening bonds between academic biobanks and industry partners.

调查目的:关于学术生物库与制药/生物技术行业之间的合作,有效合作的标准仍不明确。行业和学术生物库的研究人员可能有不同的激励机制和要求,而对方往往并不熟悉这些激励机制和要求。本调查旨在加深对这些基本知识差距的了解,这些差距可能会阻碍学术界和产业界之间的最佳合作。调查的主要结果:共有 53 位受访者。虽然这是一项全球性调查,但大多数受访者(n = 29)来自北美,这可能反映了该地区在研究方面的总体投资情况,也可能反映了学术界与产业界之间互动的增加。大多数受访的学术生物库收集多种类型的样本,其中大多数(超过 90%)同时收集生物流体(包括血液)和组织。大多数参与调查的学术生物库都意识到,他们没有(35%)或仅部分(35%)利用其库存的全部潜力。提高利用率的一个办法是与行业伙伴合作。与业界合作时遇到的主要问题被认为是各种挑战的组合,包括合同(55%)、同意限制(45%)、时限(41%)或时间压力(36%)。达成协议所需的时间也是一大障碍(54%),此外还有行业的行政要求(36%)。调查简要结论:要利用联合协作的机会,有关各方必须建立信任。第一步是了解对方的不同要求和需求,并建立高效的联合合作结构。本次调查强调了下一步加强学术生物库与行业合作伙伴之间联系的关键领域。
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引用次数: 0
Studies on the Quality and Antioxidant Status of Cryopreserved Pantja Buck Semen Supplemented with Aqueous Extract of Rosemary and Sericin in Tris Extender. 冷冻保存的 Pantja 公鹿精液中添加迷迭香水提取物和丝胶的质量和抗氧化状态研究。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2024-11-26 DOI: 10.1089/bio.2024.0082
Sunil Kumar, H P Gupta, Shiv Prasad, T K Ambwani, R K Sharma, J L Singh

Introduction: Rosemary shrub/plant and Sericin have been documented to show antioxdative properties, however their role in improving post thaw semen quality has not been well established. Objectives: The present study was conducted to investigate the effect of Rosemary leaves extract and Sericin protein on post-thaw quality of Pantja buck semen. Methods: In the first experiment, 32 ejaculates were collected from 4 sexually mature Pantja bucks and pooled to form 8 pooled samples. The pooled samples were evaluated for seminal attributes (sperm motility, viability, morphology, plasma membrane integrity, and acrosomal integrity), status of antioxidative enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) and lipid-peroxidation (malondialdehyde) of sperm plasma membrane before dilution. In the second experiment, pooled samples were divided into three equal aliquots as group-C (Control), group-R (Rosemary), and group-S (Sericin). Aliquots of group-C were diluted in glycerolated Egg Yolk Tris (EYT) extender, whereas aliquots of group-R and group-S were additionally supplemented with 4.0% v/v Rosemary leaves extract and 0.25% w/v Sericin, respectively, and again examined for the above parameters at the post-dilution, post-equilibration, and post-thawing stages of semen freezing. Results: Significant differences (p < 0.05) were observed in the values of seminal attributes, level of enzymatic antioxidants, and lipid per-oxidation between group C and R and between group C and S at the post-thaw stage of semen freezing. However Sericin was nonsignificantly better than Rosemary in improving post-thaw semen quality. Conclusion: The results of the present study on limited semen samples in Pantja buck demonstrated that compared to the control group, both Rosemary aqueous extract (4%) and Sericin protein (0.25%) as an additive in TRIS extender, showed better cryoprotective effects resulting in improved post-thaw seminal characteristics.

简介:据记载,迷迭香灌木/植物和丝胶具有抗氧化特性,但它们在改善解冻后精液质量方面的作用尚未得到充分证实。研究目的:本研究旨在探讨迷迭香灌木/植物和丝胶在改善解冻后精液质量方面的作用:本研究旨在探讨迷迭香叶提取物和丝胶蛋白对 Pantja 公鹿精液解冻后质量的影响。研究方法在第一项实验中,从 4 头性成熟的 Pantja 公鹿身上采集了 32 滴精液,并将其汇集成 8 份汇集样本。在稀释前,对混合样本进行精液属性(精子活力、存活率、形态、质膜完整性和顶体完整性)、抗氧化酶(超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和谷胱甘肽还原酶)和精子质膜脂质过氧化反应(丙二醛)的评估。在第二次实验中,将汇集的样本分成三个等量的等分组,分别为 C 组(对照组)、R 组(迷迭香组)和 S 组(丝氨酸组)。C组样本在甘油蛋黄三酸酯(EYT)扩展液中稀释,而R组和S组样本则分别添加了4.0% v/v 迷迭香叶提取物和0.25% w/v 丝裂霉素,并在精液稀释后、校准后和解冻后阶段再次检测上述参数。结果在精液冷冻后的解冻阶段,C 组和 R 组之间以及 C 组和 S 组之间的精液属性值、酶抗氧化剂水平和脂质过氧化物水平存在显著差异(p < 0.05)。然而,在改善解冻后精液质量方面,丝胶素的效果并不明显优于迷迭香。结论本研究对 Pantja 公鹿的有限精液样本进行了研究,结果表明,与对照组相比,迷迭香水提取物(4%)和丝胶蛋白(0.25%)作为 TRIS 扩展剂的添加剂,具有更好的低温保护效果,从而改善了精液的解冻后特性。
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引用次数: 0
The Evolution of Conservation Biobanking: A Literature Review and Analysis of Terminology, Taxa, Location, and Strategy of Wildlife Biobanks Over Time. 保护生物银行的演变:野生动物生物银行的术语、分类群、位置和策略的文献回顾和分析。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2025-02-12 DOI: 10.1089/bio.2024.0151
Devin M Chen, Gabriela F Mastromonaco

Nearly one-third of flora, fauna, and funga species on Earth are threatened with extinction. In response, the prevalence of repositories-often called "biobanks" or "genome resource banks"-for storing biological materials from threatened species has become more widespread. This research examined trends for the (1) terminology, (2) taxa representation, (3) global distribution, and (4) operational approach of biobanks versus genome resource banks relating to zoos and wildlife. Our literature search results indicate that although genome resource banking literature began earlier in the 1990s, biobanking has seen a surge in publications with over 3.5× more literature for biobanking since 2020. Genome resource bank articles were highly focused on mammals (68%), while biobanking literature focused more on multi-taxonomic overviews and less-studied taxa. Our search parameters found the largest number of wildlife biobanks in Europe (18) and the lowest number in South America (2), though results are likely impacted by the search being completed in English. Additionally, only 28% (7/25) of global biodiversity hotspots contain a wildlife biobank based on our methodology. While not all wildlife biobanking efforts are published or reported, these findings suggest that (1) "biobank" will likely be the more widely used term in the future, (2) more biobanking research is needed for non-mammalian taxa, (3) there are geographical gaps in wildlife biobanks, and (4) conservation biobanking programs should focus on storing biospecimens from a wide set of individuals and develop assisted reproductive technologies concomitantly with the goal of maintaining healthy, sustainable populations in the long term.

地球上近三分之一的动植物和真菌物种面临灭绝的威胁。作为回应,通常被称为“生物银行”或“基因组资源库”的储存濒危物种生物材料的储存库已经变得更加普遍。本研究考察了生物库与动物园和野生动物基因组资源库在以下方面的发展趋势:(1)术语,(2)分类群代表,(3)全球分布,以及(4)操作方法。我们的文献检索结果表明,尽管基因组资源库文献早在20世纪90年代就开始了,但自2020年以来,生物银行的出版物数量激增,生物银行的文献数量增加了3.5倍以上。基因组资源库文章高度关注哺乳动物(68%),而生物资源库文献更多地关注多分类概述和较少研究的分类群。我们的搜索参数发现,欧洲野生动物生物银行的数量最多(18个),南美洲的数量最少(2个),尽管结果可能受到英文搜索完成的影响。此外,根据我们的方法,只有28%(7/25)的全球生物多样性热点地区包含野生动物生物库。虽然并不是所有的野生动物生物银行的努力都被发表或报道过,但这些发现表明:(1)“生物银行”可能会成为未来更广泛使用的术语,(2)需要对非哺乳动物分类群进行更多的生物银行研究,(3)野生动物生物银行存在地理差距,(4)保护生物银行计划应侧重于储存来自广泛个体的生物标本,并开发辅助生殖技术,以长期维持健康,可持续的种群。
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引用次数: 0
Seminal Plasma-Derived Exosome Preserves the Quality Parameters of the Post-Thaw Semen of Bulls with Low Freezeability. 精浆衍生外泌体保存低冷冻性公牛解冻后精液的质量参数。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2024-12-26 DOI: 10.1089/bio.2024.0077
Rahele Ranjbar Shamsi, Razi Jafari Jozani, Reza Asadpour, Maryam Rahbar, Morteza Taravat

Introduction: Sperm cryopreservation is a useful storage technique in artificial insemination. Nanoparticles and nanovesicles such as exosomes are widely used in sperm cryopreservation procedures to alleviate cold-induced injury inflicted during sperm freezing. Objective: The objective of the present study was to examine the impact of varying concentrations of exosomes derived from seminal plasma added to a freezing extender on the quality of post-thawed bull sperm. Methods: Five Holstein bulls were chosen based on their samples having less than 30% progressive motility. After exosome extraction, semen samples from bulls (n = 5) with progressive sperm motility ≤30% were collected, diluted with different exosome concentrations (0, 25, 50, and 100 μg/mL), and aspirated into 0.5 mL straws. After the freeze-thaw process, sperm total and progressive motility, viability, morphology, plasma membrane integrity, mitochondrial activity, and apoptosis status were assessed. Furthermore, the expression levels of annexin (ANX1), dystrophy-associated Fer-1-like protein (DYSF), fibronectin 1 (FN1), and reactive oxygen species modulator 1 (ROMO1) were evaluated via real-time polymerase chain reaction (PCR). Results: Adding different concentrations of exosomes (25, 50, and 150 μg/mL) significantly increased the progressive motility, viability, and membrane integrity of sperm compared with the control group (p < 0.05). For the apoptosis index, treatment with 100 μg/mL exosomes significantly increased the percentage of live cells (p < 0.05), while the percentage of necrotic cells decreased significantly (p < 0.05) compared with 25 μg/mL exosome. The results of quantitative PCR showed that the expression levels of ANX1 were significantly (p < 0.05) upregulated at 50 μg/mL exosome, and the expression of ROMO1, FN1, and DYSF were downregulated upon treatment with different exosome concentrations. Conclusions: In conclusion, supplementing the freezing diluent with exosome-derived seminal plasma could preserve the quality parameters of the post-thaw semen of the bull with low freezeability and could be used as a helpful method for reproductive programs.

精子低温保存是人工授精的一种有效的保存技术。纳米粒子和纳米囊泡(如外泌体)被广泛应用于精子冷冻保存过程中,以减轻精子冷冻过程中造成的冷损伤。目的:本研究的目的是研究从精浆中提取的不同浓度的外泌体加入冷冻扩展剂对解冻后公牛精子质量的影响。方法:选取5头进行性运动性低于30%的荷斯坦公牛为研究对象。提取外泌体后,收集精子活力≤30%进行性公牛精液样本(n = 5),用不同浓度的外泌体(0、25、50、100 μg/mL)稀释后,抽吸至0.5 mL吸管中。在冻融过程后,评估精子的总活力和进展活力、活力、形态、质膜完整性、线粒体活性和凋亡状态。此外,通过实时聚合酶链反应(PCR)评估膜联蛋白(ANX1)、营养不良相关的fe -1样蛋白(DYSF)、纤维连接蛋白1 (FN1)和活性氧调节剂1 (ROMO1)的表达水平。结果:与对照组相比,添加不同浓度的外泌体(25、50、150 μg/mL)可显著提高精子的进行活力、活力和膜完整性(p < 0.05)。在细胞凋亡指数方面,与25 μg/mL外泌体相比,100 μg/mL外泌体显著提高了活细胞百分比(p < 0.05),坏死细胞百分比显著降低(p < 0.05)。定量PCR结果显示,在50 μg/mL外泌体处理下,ANX1的表达水平显著上调(p < 0.05), ROMO1、FN1和DYSF的表达在不同浓度外泌体处理下均下调。结论:外泌体源性精浆添加冷冻稀释液可以保存低冷冻性公牛解冻后精液的质量参数,可作为一种有益的生殖方案。
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引用次数: 0
Standard PREanalytical Code Version 4.0. 标准预分析代码 4.0 版。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2024-08-12 DOI: 10.1089/bio.2024.0010
Fay Betsou, Rodrigo Chuaqui, Annemieke De-Wilde, William Grizzle, Michael Kiehntopf, Loes Linsen, William Mathieson, Ioana Moldovan, Rocío Aguilar-Quesada, Katheryn Shea, Purnima Singh, Stella Somiari, Demetri Spyropoulos, Juhi Tayal, Riad Tebbakha, Klara Valyi-Nagy, Lalita Wadhwa
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引用次数: 0
Trehalose Cryopreservation of Human Mesenchymal Stem Cells from Cord Tissue. 人脐带间充质干细胞的海藻糖低温保存。
IF 1.4 4区 生物学 Pub Date : 2025-08-01 Epub Date: 2024-12-26 DOI: 10.1089/bio.2024.0025
Nuria Izaguirre-Pérez, Gertrudis Ligero, Paula Alba Aguilar-Solana, José Antonio Carrillo-Ávila, Carmen Ruth Rodriguez-Reyes, Ida Biunno, Rocío Aguilar-Quesada, Purificación Catalina

Adequate hypothermic storage of human mesenchymal stem cells (hMSCs) is of fundamental importance since they have been explored in several regenerative medicine initiatives. However, the actual clinical application of hMSCs necessitates hypothermic storage for long periods, a process that requires the use of non-toxic and efficient cryo-reagents capable of maintaining high viability and differentiating properties after thawing. Current cryopreservation methods are based on cryoprotectant agents (CPAs) containing dimethylsulphoxide (DMSO), which have been shown to be toxic for clinical applications. In this study, we describe a simple and effective trehalose (TRE)-based solution to cryo-store human umbilical cord-derived MSCs (UC-MSCs) in liquid nitrogen. Cells viability, identity, chromosomal stability, proliferative and migration capacity, and stress response were assessed after cryopreservation in TRE as CPA, testing different concentrations by itself or in combination with ethylene glycol (EG). Here we show that TRE-stored UC-MSCs provided lower cell recovery rates compared with DMSO-based solution, but maintained good functional properties, stability, and differentiating potential. The best cell recovery was obtained using 0.5 M TRE with 10% EG showing no differences in the osteogenic, adipogenic, and chondrogenic differentiation capacity. A second cycle of cryopreservation in this TRE-based solution had no additional impact on the viability and morphology, although slightly affected cell migration. Furthermore, the expression of the stress-related genes, HSPA1A, SOD2, TP53, BCL-2, and BAX, did not show a higher response in UC-MSCs cryopreserved in 0.5 M TRE + 10% EG compared with DMSO. Together these results, in addition to ascertained therapeutic properties of TRE, provide sufficient evidence to consider TRE-based medium as a low-cost and efficient solution for the storage of human UC-MSCs cells and potentially substitute DMSO-based cryo-reagents.

人间充质干细胞(hMSCs)的充分低温储存是至关重要的,因为它们已经在几个再生医学计划中进行了探索。然而,hMSCs的实际临床应用需要长期低温储存,这一过程需要使用无毒高效的冷冻试剂,能够在解冻后保持高活力和分化特性。目前的冷冻保存方法是基于含有二甲基亚砜(DMSO)的冷冻保护剂(cpa),这在临床应用中已被证明是有毒的。在这项研究中,我们描述了一种简单有效的基于海藻糖(TRE)的解决方案,用于在液氮中冷冻储存人脐带来源的间充质干细胞(UC-MSCs)。通过单独或与乙二醇(EG)联合使用不同浓度的TRE冷冻保存后,评估细胞活力、身份、染色体稳定性、增殖和迁移能力以及应激反应。本研究表明,与基于dmso的溶液相比,tre3储存的UC-MSCs提供了较低的细胞回收率,但保持了良好的功能特性、稳定性和分化潜力。使用0.5 M TRE和10% EG获得最佳细胞回收率,在成骨、成脂和成软骨分化能力方面没有差异。在这种基于trec的溶液中进行第二轮冷冻保存对细胞的活力和形态没有额外的影响,尽管对细胞迁移有轻微的影响。此外,与DMSO相比,在0.5 M TRE + 10% EG中冷冻保存的UC-MSCs中,应激相关基因HSPA1A、SOD2、TP53、BCL-2和BAX的表达并没有表现出更高的反应。综上所述,除了确定了TRE的治疗特性外,这些结果提供了足够的证据,证明基于TRE的培养基是一种低成本、高效的储存人UC-MSCs细胞的解决方案,并有可能取代基于dmso的冷冻试剂。
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引用次数: 0
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Biopreservation and Biobanking
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