Sato N, Fricke C, McGuckin C, Forraz N, Degoul O, Atzeni G, Sakurai H
Medical Devices Division, Kaneka Corporation, Osaka, 530-8288, Japan.
Kaneka Pharma Europe N.V. German Branch, DE-65760, Eschborn, Germany.
Cell Prolif. 2015 Dec;48(6):671-81. doi: 10.1111/cpr.12217. Epub 2015 Oct 12.
Availability of cord blood (CB) processing has been limited by the need for electrically aided centrifugal techniques, which often produce only low final cell product yield. Here, we describe development and characterization of a novel filter device aimed at allowing CB processing, using gentle gravity-led flow.
CB was processed with a novel filter device (CellEffic CB, consisting of non-woven fabric), without any centrifugation. Cells were harvested by flushing the filter with either HES or physiological saline solution (SALINE). Differential cell counts and viability analysis, combined with Fluorescence-Activated Cell Sorting (FACS) (total nucleated cells [TNC], mononuclear cells [MNC], CD45+ CD34+ cells, hematopoietic precursor cells [HPCs]) and clonogenic assay, were employed for analysis of CB pre- and post-processing, and after freeze/thawing.
Processing using the novel filter yielded high quality RBC depletion while maintaining good recovery of TNC, MNC, CD34+, HPCs and colony forming unit (CFU) output. The filter performed equally well using HES or SALINE. Gravity-led flow provided gentle cell movement and protection of the stem cell compartment. Post-thaw CFU output was maintained particularly, an important indicator for CB banking.
Geographical limitations of CB transplantation and banking have required a non-electrical, non-centrifugal solution. This novel filter CellEffic CB device revealed rapid yet gentle cell processing while maintaining the stem/progenitor cell compartment required for both haematological and regenerative medicine therapies.
脐带血(CB)处理因需要电动辅助离心技术而受到限制,这种技术通常只能产生较低的最终细胞产品产量。在此,我们描述了一种新型过滤装置的开发和特性,该装置旨在通过温和的重力引导流来实现CB处理。
使用一种新型过滤装置(CellEffic CB,由无纺布组成)处理CB,无需任何离心操作。通过用羟乙基淀粉(HES)或生理盐水(SALINE)冲洗过滤器来收获细胞。采用细胞分类计数和活力分析,结合荧光激活细胞分选(FACS)(总核细胞 [TNC]、单核细胞 [MNC]、CD45+ CD34+ 细胞、造血前体细胞 [HPC])和克隆形成试验,对CB处理前、处理后以及冻融后的情况进行分析。
使用新型过滤器进行处理可实现高质量的红细胞去除,同时保持TNC、MNC、CD34+、HPC和集落形成单位(CFU)产量的良好回收率。使用HES或SALINE时,该过滤器的性能相同。重力引导流使细胞移动温和,并保护干细胞区室。解冻后的CFU产量尤其得以维持,这是CB储存的一个重要指标。
CB移植和储存的地理限制需要一种非电动、非离心的解决方案。这种新型过滤器CellEffic CB装置在保持血液学和再生医学治疗所需的干/祖细胞区室的同时,实现了快速而温和的细胞处理。