A High Performance Point-of-Care Device for Umbilical Cord Blood Processing

描述(由申请人提供)：该项目旨在继续开发和验证一种在第一阶段开发的新型微流控设备，该设备可以最大限度地保持脐带血造血干细胞(HSC)的活性、回收率和功能。异基因造血干细胞移植是包括白血病和淋巴瘤在内的50多种危及生命的疾病的选择性治疗。然而，由于捐赠者的匹配，只有不到30%的患者实际获得了捐赠者。脐带血已经成为一种容易获得的HSC来源，因为它已经被收集并储存在世界各地的脐带血库中。然而，今天储存的单位中可存活的造血干细胞的数量不足以有效地治疗成年患者。为了将成人患者的一年存活率从目前的~20%显著提高到儿童患者的50%-70%的成功率，估计移植的脐带血单位中存活的HSC数量必须增加~3倍。研究表明，目前的脐带血加工和储存做法导致HSC产量和功能损失高达90%，特别是在运输延迟(36-48小时)和离心量减少步骤期间，只有10-20%的原始HSC群体用于治疗。我们在第一阶段开发的新设备能够在护理地点简单、即时地处理脐带血，只需最少的设备和人员培训，从而将生存能力和产量的损失降至最低。第一阶段的原型已经证明了使用微流控技术处理整个绳索单元的可行性，并且比目前的行业标准要好得多-- CD34+HSC的回收率为94%，无活性丧失或体外造血祖细胞活性丧失(n=12)。第二阶段的目标是：1.开发优化的工作原型，以当前脐带血库中使用的领先的基于离心机的系统为基准，验证性能和基准。2.在临床环境中演示护理点的设备实用性。3.确保设备柔和 和生物惰性，通过使用流式细胞仪免疫表型来表征细胞激活。4.通过移植到免疫缺陷的人源化小鼠模型中，显著提高了回收细胞在体内的植入能力。该项目的潜在影响包括使脐带血成为移植HSC的主要来源，为成年患者提供明显更好的结果，以及使脐血库和基于HSC的治疗更有效。

DESCRIPTION (provided by applicant): This project aims to continue development and validation of a novel microfluidics device developed during Phase I that preserves the maximal viability, recovery, and function of cord blood hematopoietic stem cells (HSC). Allogeneic HSC transplantation is the elective treatment for over 50 life threatening diseases including leukemia and lymphoma. However due to matching donor availability, fewer than 30% of the patients actually receive one. Umbilical cord blood has emerged as a readily available source of HSC as it has been collected and stored in cord blood banks worldwide. However, the number of viable HSC in the units stored today is insufficient to effectively treat adult patients. For the 1yr survval rate of adult patients to be significantly increased from the current ~20% to the 50% - 70% success rate achieved in children, it is estimated that the number of viable HSC in a transplanted cord blood unit has to be increased by a factor of ~3. Studies have shown that the current cord blood processing and banking practices lead to as much as ~90% loss in HSC yield and function, particularly during the shipping delay (36 to 48 hrs) and the centrifugal volume reduction step, leaving only 10 - 20% of the original HSC population for treatment. The new device we developed during Phase I enables simple, immediate processing of cord blood at point of care with minimal equipment and training of personnel, thereby minimizing the loss in viability and yield. Phase I prototypes have demonstrated feasibility of using microfluidics to process entire units of cord and significantly better results than the current industry standards-- ~94% of the CD34+ HSC were recovered with no loss of viability or in vitro progenitor activity (n =12). Phase II aims to: 1. Develop optimized working prototype, validate performance and benchmark against leading centrifugation-based systems used in current cord blood banks. 2. Demonstrate device utility at the point of care in a clinical setting. 3. Ensure device gentleness and biological inertness by characterizing cell activation using flow cytometric immune-phenotyping. 4. Demonstrate significant improvement of in vivo engraftment capacity of recovered cells by engraftment into immune-deficient humanized mouse models. Potential impacts of the project include enabling cord blood to become the primary source of HSC for transplantation, delivering significantly better outcomes for adult patients, and making cord blood banking and HSC-based therapies more effective.