AUTOMATED CLOSED SYSTEM FOR DMSO REMOVAL FROM PERIPHERAL BLOOD STEM CELLS

描述（由申请人提供）：大剂量化疗后外周血造血干细胞（PBSC）移植常用于治疗各种血液恶性肿瘤。为了促进这一过程，通常收集自体PBSC，冷冻保存并保存一段时间。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，冷冻解冻的细胞与DMSO一起移植回患者体内。然而，长期以来，DMSO回输与各种不良事件相关，从皮肤潮红、头痛、发烧、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系统事件。这些可能导致发病率增加、住院时间延长和治疗相关费用增加。目前，当试图去除DMSO时，通常需要使用离心机对细胞进行“清洗”。这些方法引入机械力和渗透应力，导致细胞堆积/结块和潜在的重大细胞损失。他们还需要额外的实验室工作人员，并且每个病人需要3到4个小时的工作，这使得许多机构的手术更加昂贵和实际困难。最后，在细胞洗涤过程中，很难将细胞保持在一个“封闭”的系统中，这可能导致污染。本提案的目标是改进和优化一种自动化的、便携式的、封闭系统的扩散驱动洗涤装置，用于清除PBSC中的DMSO，以防止在临床移植过程中向患者输注DMSO而不损失细胞数量或细胞功能。在这项I期研究中，该设备将被优化，以有效清除细胞悬液中的DMSO，同时通过体外克隆生成能力和植入NOD/SCID小鼠模型来评估细胞的高回收率和维持细胞功能活力。

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.