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.

项目概要 该提案的总体目标是推进干细胞生物学、基于细胞的诊断和细胞学领域的发展。 使用系统分离特定干细胞的新途径采购细胞治疗和药物开发 我们提到的肌肉骨骼结缔组织中多个生态位的细胞和祖细胞亚型 我们的目标是为患者、鸡群和研究人员提供“基于绩效的选择”(PBS)。 他们需要的知识以及严格且可重复的工具和标准，以确定并获得最佳的 细胞可用于再生和关节保存疗法、药物发现和基础研究。 间充质基质细胞 (MSC) 源自于建立结缔组织祖细胞 (CTP) 的集落 骨骼和其他组织对于细胞治疗具有广泛的价值，然而，批次之间的 MSC 存在差异。 代表了生物医学研究界持续存在的一个深刻的“痛苦”，限制了临床研究 基于 MSC 的产品的临床治疗需要高水平的可重复性和性能。 最大限度地减少变异对于推进细胞疗法的新兴领域至关重要。 我们发现，这种变化的一个重要根源是基于 MSC 制造的传统方法的失败。 在“竞争性扩张”（CE）上，用户只需将组织来源的所有细胞放入培养物中，在那里它们 对于要包含或排除的单元格没有做出任何选择，用户只是接受可能的结果。 通过初始培养物中集落形成细胞的类型和混合的随机变化来预先确定。 该提案集成了多个独特的技术平台，以实现快速有效的进展 克服这些限制，具体来说： 1) 用于自动化大视场 (LFOV)（“Google 地球”）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) 延时视频显微镜能够连续跟踪每个 CTP 衍生集落的形成 从电镀时开始，倒转视频使我们第一次能够看到并描述菌落的特征。 建立CTP，因为它们从我们的身体中取出后、分裂前立即存在。 3) 自动化机器人平台 (Cell X™)，可以选择（“挑选”）单个 CTP 衍生品 克隆以及“解放双手”扩展其后代所需的自动化处理，使 与当前的手动方法相比，精度和重现性大大提高。 该提案建立了 CTP 的生物潜力差异很大的知识，我们将使用这些工具。 确定应该或不应该包含在 MSC 制造中的集落建立 CTP 的属性 然后提供使用这些知识来实现​​高质量和高度可重复的结果的方法。