A NOVEL FROZEN STORAGE SYSTEM FOR CORD BLOOD STEM CELLS

一种新型脐带血干细胞冷冻储存系统

• 批准号: 6698852
• 负责人: ERIK J. WOODS
• 金额: $ 49.88万
• 依托单位: GENERAL BIOTECHNOLOGY, LLC
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6698852
• 关键词: NOD mouse; SCID mouse; blood banks; cord blood; cryopreservation; dimethylsulfoxide; freezing; hematopoietic stem cells; human subject; mathematical model; placenta; technology /technique development; tissue /cell preparation; tissue resource /registry

DESCRIPTION (provided by applicant): With increasing effort directed toward purification and/or in vitro expansion of hematopoietic progenitor cells (HPCs) for either fundamental research or clinical applications, there is an associated, increased need for the development of improved, efficient preservation methods for these cells to provide "cell banks." In addition, with the increase in allograft of purified progenitor cells, the possibility of disease transmission from use of contaminated donor material requires that only cells from infectious disease-screened donors be used. The ability to store frozen cells would allow sufficient time for adequate screening to be performed. Therefore, the overall goal of this continuing research is to further develop and optimize an improved system for cryopreservation and storage of human placental/umbilical cord blood (PCB) derived hematopoietic progenitor cells (HPCs), especially as it relates to improving the retention of normal structure and function of frozen-thawed cells to improve clinical outcome (engraftment). In Phase I, a system was developed which provides (1) an efficient, simplified method of bulk-freezing HPC units, and (2) optimized removal of cryoprotectant (DMSO) while maintaining a closed system. In Phase II, this system will be refined to allow (1) ease of use in the clinical environment, and (2) ease of manufacture/commercialization. It is the goal of this Phase II proposal to refine the closed cryopreservation, storage, and PCB washing system based on sound biophysical cell-type specific characteristics and mathematical modeling which will allow efficient and safe use of cryopreserved PCB derived HPCs. To achieve this goal, the following specific aims are proposed: (1) Validation and refinement of the cryopreservation method/system-using colony forming assays in semi-solid media to determine clonogenic ability, (2) Validation and refinement using a human long-term culture initiating cell (LTC-IC) assay, and (3) Testing of the cryopreservation system by freezing/thawing/washing HPCs and transplanting into a NOD/SCID mouse model.

描述（由申请人提供）：随着针对用于基础研究或临床应用的造血祖细胞（HPC）的纯化和/或体外扩增的努力的增加，对开发用于这些细胞的改进的、有效的保存方法以提供“细胞库”的相关的、增加的需求。“此外，随着纯化祖细胞同种异体移植的增加，使用受污染的供体材料传播疾病的可能性要求只使用来自感染性疾病筛查供体的细胞。储存冷冻细胞的能力将允许有足够的时间进行适当的筛选。因此，该持续研究的总体目标是进一步开发和优化用于冷冻保存和储存人胎盘/脐带血（PCB）衍生的造血祖细胞（HPC）的改进系统，特别是因为其涉及改善冻融细胞的正常结构和功能的保留以改善临床结果（植入）。在第I阶段，开发了一种系统，该系统提供（1）批量冷冻HPC单元的有效简化方法，以及（2）在保持封闭系统的同时优化冷冻保护剂（DMSO）的去除。在第II阶段，将对该系统进行改进，以实现（1）在临床环境中易于使用，以及（2）易于制造/商业化。本II期提案的目标是基于合理的生物物理细胞类型特异性特征和数学建模来完善封闭式冷冻保存、储存和PCB洗涤系统，这将允许高效和安全地使用冷冻保存的PCB衍生HPC。为实现这一目标，提出了以下具体目标：（1）冷冻保存方法/系统的验证和改进-使用半固体培养基中的集落形成测定来确定克隆形成能力，（2）使用人长期培养起始细胞（LTC-IC）测定的验证和改进，和（3）通过冷冻/解冻/洗涤HPC并移植到NOD/SCID小鼠模型中来测试冷冻保存系统。