AUTOMATED CLOSED SYSTEM FOR DMSO REMOVAL FROM PERIPHERAL BLOOD STEM CELLS

描述(申请人提供)：大剂量化疗后，外周血造血干细胞(PBSC)移植是治疗各种血液系统恶性肿瘤的常用方法。为了促进这一过程，通常收集、冷冻保存自体PBSC并储存一段时间。目前的PBSC冷冻保存需要使用摩尔浓度的冷冻保护剂添加剂二甲基亚砜(DMSO)。通常情况下，冷冻解冻的细胞会与DMSO一起移植回患者体内。然而，回输DMSO长期以来一直与各种不良事件有关，从皮肤潮红、头痛、发热、呼吸困难、腹部痉挛、恶心和腹泻，到更严重的影响，如溶血、心血管症状和脑血管缺血导致神经事件。这可能会导致发病率增加、住院时间延长和与治疗相关的费用增加。目前，当尝试去除DMSO时，程序通常涉及使用离心法对细胞进行“清洗”。这些方法引入机械力和渗透压力，导致细胞堆积/聚集和潜在的显着细胞损失。他们还需要额外的实验室工作人员，每个患者需要3到4个小时的工作，这使得这一过程对许多机构来说更加昂贵和困难。最后，在细胞清洗过程中，很难将细胞保持在封闭的系统中，这可能会导致污染。这项建议的目标是改进和优化一种自动化、便携式、闭合系统扩散驱动的洗涤设备，用于从PBSC中清除DMSO，以防止在临床移植期间向患者输注DMSO，而不会损失细胞数量或细胞功能。在这项第一阶段的研究中，该设备将进行优化，以有效地清除细胞悬浮液中的二甲基亚砜，同时允许高细胞回收率，并通过体外克隆形成能力和NOD/SCID小鼠模型的植入来维持细胞功能活性。

DESCRIPTION (provided by applicant): High dose chemotherapy followed by peripheral blood hematopoietic stem cell (PBSC) transplantation is frequently used for treating various hematologic malignancies. To facilitate this process, autologous PBSC are typically collected, cryopreserved and stored for some period of time. Current PBSC cryopreservation requires the use of molar concentrations of the cryoprotectant additive dimethyl sulfoxide (DMSO). Typically, frozen-thawed cells are transplanted back into patients along with DMSO. However, reinfusion of DMSO has long been associated with various adverse events, ranging from skin flushing, headache, fever, dyspnea, abdominal cramping, nausea, and diarrhea to more sever effects such as, hemolysis, cardiovascular symptoms and cerebrovascular ischemia leading to neurological events. These can result in increased morbidity, prolonged hospitalization, and increased treatment-related costs. Currently, when removal of DMSO is attempted, the procedures typically involve cell "washing" using centrifugation. These methods introduce mechanical forces and osmotic stress causing cell packing/clumping and potential significant cell loss. They also require additional laboratory staff as well, and take between 3 and 4 hours of work per patient, which makes the procedure more expensive and practically difficult for many institutions. Finally, it is difficult to keep the cells in a "closed" system during cell washing, which may lead to contamination. It is the goal of this proposal to refine and optimize an automated, portable, closed-system diffusion-driven washing device for use in clearance of DMSO from PBSC, to prevent infusion of DMSO into patients during clinical transplant without loss of cell numbers or cell functionality. In this Phase I research, the device will be optimized for effective clearance of DMSO from cell suspension solutions while allowing high cell recovery with maintenance of cell functional viability as assessed by in vitro clonogenic ability as well as engraftment in a NOD/SCID mouse model.