Optimizing Industrial-Scale GMP Processes for Recovering and Banking Deceased Donor Bone Marrow

摘要 本项目旨在优化一种从器官中储存临床级骨髓的方法。 可以在全国范围内部署以缓解短缺的捐赠者，提供骨髓储备 作为应对核威胁的医学对策，并支持现有的研究方法，如 作为延迟免疫耐受诱导和输注艾滋病毒耐药干细胞。 在第一阶段，我们采用了由我们的研究团队开发的协议，用于采购、处理、 和储存来自身体椎体和髂骨的骨髓，这被证明产生了 供者移植后活的造血干细胞和其他骨髓细胞的高产量 不同长度的热缺血和冷缺血时间。根据这些数据推导出了方程式 从而可以对热缺血和冷缺血对造血干细胞的影响进行建模 生存能力和功能。这些预测模型提供了完成以下任务所需的工具 回收和隔离方案的优化。在这项提案中，我们将进行关键的改进 到我们的大规模部署方法，这将允许在不同的地理位置进行恢复 器官采购机构和运输到一个中央设施进行处理。最终的结果是 将有大量的骨髓供应，可以按需使用，现成，补充 现有的骨髓捐赠者登记和脐带血库。 这项第二阶段研究的技术目标是(1)优化干细胞在 整块脊椎被运往铯的中央处理设施，(2)最大限度地回收可存活的 干细胞暴露在长时间的热缺血中，以及(3)验证最终优化的工艺。在……里面 目标1，我们将优化供体椎体采购和运输的保存方案 身体骨髓。在目标2中，我们将优化整个骨髓分离方案，以最大限度地 活细胞的恢复。在目标3中，我们将通过建立和测试一家试点银行来验证所有流程 在小鼠异种移植中比较已故供者和抽出的骨髓 学习。这些目标的结果将决定最有效的采购制度， 身体骨髓的加工和储存，以供临床大规模使用。

ABSTRACT This program aims to optimize a method for banking clinical-grade bone marrow from organ donors that can be deployed nationally to ameliorate shortages, provide bone marrow stockpiles as medical countermeasures for nuclear threats, and support existing research approaches such as delayed immune tolerance induction and transfusion of HIV-resistant stem cells. In Phase I we adapted protocols developed by our research team for procurement, processing, and banking bone marrow from cadaveric vertebral bodies and ilia, which were shown to produce high yields of viable hematopoietic stem cells and other bone marrow cells from donors after varying lengths of warm and cold ischemic times. Equations have been derived from these data which have allowed modeling the effects of warm and cold ischemia on hematopoietic stem cell viability and function. These predictive models provide the tools necessary for completing optimization of the recovery and isolation protocols. In this proposal we will make key refinements to our method for deployment on a large scale, which will allow recovery at geographically diverse organ procurement agencies and shipment to a centralized facility for processing. The end result will be a large supply of bone marrow that can be used on-demand, off the shelf, complementing existing bone marrow donor registries and cord blood banks. The technical objectives of this Phase 2 study are to (1) optimize preservation of stem cells in whole vertebrae shipped to Ossium’s central processing facility, (2) maximize recovery of viable stem cells exposed to long warm ischemia times, and (3) validate the final optimized process. In Aim 1, we will optimize preservation solutions for procurement and shipment of donor vertebral body bone marrow. In Aim 2, we will optimize whole bone marrow isolation solutions to maximize recovery of viable cells. In Aim 3, we will validate all processes by building and testing a pilot bank and comparing deceased donor and aspirated bone marrow in a mouse xenotransplantation study. The outcome of these aims will determine the most effective system for procurement, processing and banking of cadaveric bone marrow for clinical use on a large scale.