Understanding and Using Variation in Source Materials for MSC Fabrication

了解和使用 MSC 制造的原材料变化

• 批准号: 10614506
• 负责人: George F Muschler
• 金额: $ 63.04万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-14 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614506
• 关键词: Acceleration; Address; Advanced Development; Animal Model; Articulation; Automation; Basic Science; Biological; Biological Assay; Biology; Biomedical Research; Cartilage; Cell Cycle Kinetics; Cell Therapy; Cell surface; Cells; Cellular Assay; Clinical; Clonal Expansion; Clone Cells; Communities; Connective Tissue; Controlled Environment; Data; Development; Diagnostic; Disease; Drug Delivery Systems; Engineering; Excision; Exclusion; Failure; Fatty acid glycerol esters; Functional disorder; Future; Goals; Hand; Health; Histologic; Human; Image; Image Analysis; In Vitro; Individual; Joints; Kinetics; Knowledge; Manuals; Marrow; Measurable; Measures; Methods; Morphology; Musculoskeletal; Outcome; Pathway interactions; Patients; Performance; Persistent pain; Planet Earth; Population; Process; Proliferating; Reproducibility; Research; Research Personnel; Resources; Robotics; Series; Source; Standardization; Surface; Testing; Time; Tissues; Transplantation; Variant; Video Microscopy; Work; automated image analysis; bone; cell type; clinical application; drug development; drug discovery; fabrication; histological specimens; imaging platform; immunoregulation; improved; in vivo; induced pluripotent stem cell; insight; manufacture; mesenchymal stromal cell; migration; novel; preservation; progenitor; regenerative; research clinical testing; scaffold; stem; stem cell biology; stem cell population; stem cells; technology platform; therapy development; tool

Project Summary The Broad Aim of this proposal is to advance the field of stem cell biology, cell-based diagnostics, and cell sourcing for cell therapy and drug development using a new pathway for systematic isolation of specific stem cell and progenitor cell subtypes from the plurality of niches in musculoskeletal connective tissues that we refer to as “Performance-Based Selection” (PBS). Our goal is to provide patients, clinicians and researchers with the knowledge and rigorous and reproducible tools and standards that they need, to identify and obtain the best possible cells for use in regenerative and joint preservation therapies, drug discovery and basic research. Mesenchymal Stromal Cells (MSCs) derived from colony founding Connective Tissue Progenitors (CTPs) in bone and other tissues are widely valuable for cellular therapy. However, batch to batch MSC variation represents a profound point of ongoing “pain” in the biomedical research community that is limiting the clinical evaluation and performance of MSC-based products. Clinical therapies demand high levels of repeatability and reproducibility. Minimizing variation is essential to advancing the emerging field of cellular therapies. We hypothesize that a large source of this variation is the failure of traditional methods of MSC fabrication based on “Competitive Expansion” (CE), where the user simply puts all cells from tissue source into culture, where they compete. No choice is made regarding cells to include or exclude. The user simply accepts an outcome that may be predetermined by random variation in the type and mix of colony founding cells in the initial culture. This proposal integrates several unique technology platforms to enable rapid and effective progress in overcoming these limitations, specifically: 1) Standardized robotics for automated large-field-of-view (LFOV) (“Google Earth”) imaging and image analysis to identify and measure CTPs and their progeny. 2) Time Lapse Videomicroscopy that enables continuous tracking of the formation of each CTP-derived colony from the time of plating. Reversing the video enables us, for the first time, to see and to characterize colony founding CTPs as they exist immediately after removal from our bodies and before they divide. 3) An automated robotic platform (Cell X™) that enables both the selection (“Picking”) of individual CTP-derived clones as well as the automated processing that is needed to expand their progeny “hands free”, enabling vastly greater precision and reproducibility than current manual methods. This proposal builds the knowledge that CTPs vary widely in biological potential. Using these tools, we will determine the attributes of colony founding CTPs that should or should not be included in MSC manufacturing and then provide the means to use this knowledge to achieve high quality and highly reproducible outcomes.

项目摘要 该建议的广泛目的是提高干细胞生物学，基于细胞的诊断和细胞的领域 使用新的途径进行细胞疗法和药物开发的采购，以系统地隔离特定的茎 来自肌肉骨骼连接的时机中的多个壁ches的细胞和祖细胞亚型，我们参考 作为“基于绩效的选择”（PBS）。我们的目标是为患者，临床医生和研究人员提供 他们需要的知识以及严格且可重复的工具和标准，以识别和获得最好的 可能用于再生和联合保存疗法，药物发现和基础研究的细胞。 源自集菌结缔组织祖细胞（CTP）的间充质基质细胞（MSC） 骨骼和其他时机对于细胞疗法而言广泛有价值。但是，批次到MSC变体 代表了生物医学研究社区中持续的“疼痛”的深刻点，这限制了临床 基于MSC的产品的评估和性能。临床疗法需要高水平的可重复性和 可重复性。最小化变异对于推进细胞疗法的新兴领域至关重要。 我们假设这种差异的很大来源是基于MSC制造的传统方法的失败 在“竞争扩展”（CE）上，用户只是将所有细胞从组织来源放入培养物中， 竞争。关于包括或排除的细胞没有任何选择。用户只是接受可能的结果 通过在初始培养物中菌落基础的类型和混合物的随机变化来预先确定。 该提案集成了几个独特的技术平台，以实现快速有效的进展 特别克服这些局限性： 1）自动化大型视野（LFOV）（“ Google Earth”）成像和图像分析的标准化机器人技术 识别和测量CTP及其后代。 2）延时视频显微镜可以连续跟踪每个CTP衍生的菌落的形成 从电镀时开始。逆转视频使我们能够第一次看到和表征殖民地 从我们的身体和分裂之前，它们立即存在，因为它们立即存在。 3）一个自动机器人平台（Cell X™），该平台可以启用单个CTP衍生的选择（“选择”） 克隆以及所需的自动处理，以扩大其进度的“免费”，从而实现 与当前的手动方法相比，精度和可重复性要高得多。 该建议建立了CTP在生物潜力方面差异很大的知识。使用这些工具，我们将 确定殖民地创始CTP的属性，该CTP应该或不应包括在MSC制造中 然后提供使用这些知识来实现​​高质量和高度再现结果的方法。