CONTINUOUS MAGNETIC CELL SORTING

根据细胞的功能或表面属性分离细胞是一种 细胞生物学、癌症治疗和生物医学的许多技术基础 生物技术。虽然免疫磁性细胞分离已被用于 所有这些领域，都受到了相对较大的 与靶细胞相比的顺磁性标记以及通过相对 用于分离的粗制设备。这项研究将有两个主要目标： 一种新型磁性细胞标签的研制及其分析和 设计了一种使用该标签的连续磁性细胞分选机。这是一项新的 磁性标记由铁蛋白与单抗结合而成 针对靶细胞上的特定表面抗原。铁蛋白是一种 哺乳动物体内的铁储存蛋白。 铁蛋白标记物的特性将使用一些 技术包括：核磁共振波谱，原子 吸收光谱和透射电子显微镜。独一无二的 结合荧光和磁性标签的三明治标签技术 与流式细胞术结合用于细胞标记的表征 有约束力的。人外周血淋巴细胞与人乳腺癌的关系 在这项研究中，细胞将与单抗结合使用。 特异性标志物抗体：CD3、CD4、CD8、CD16、CD34和 伯尔-第四集。这些标志物识别对骨髓重要的细胞。 移植治疗：T细胞，辅助/诱导者，细胞毒/抑制者， 自然杀伤细胞、干细胞和乳腺癌细胞。 由于所涉及的复杂性，分析和设计一种连续的、 磁性细胞分选机将需要使用三维粒子图像 血流速度测定仪(3D PIV)。由俄亥俄州立大学开发的3D PIV， 这项研究的拟议分包地点将允许确定 强磁场中标记细胞和未标记细胞三维速度矢量的比较 菲尔兹。通过实验分析，结合相关方程，给出了计算结果 Motion，将提供微米级的系统信息。使用 这些信息将被开发出一种优化的细胞分类器。此外, 将有可能设计更先进的分选器，如多级 可以获得非常高纯度或非常高选择性的系统 在相对较短的时间内。 最终目标是开发一种连续的、磁性的细胞分选器 骨髓移植治疗中的大规模分离。这样的一个 分选机将适用于快速、大规模的T细胞耗尽、癌症 细胞净化和造血干细胞浓缩。

Separation of cells based on their function or surface properties is a basis of many techniques used in cell biology, cancer therapy and in biotechnology. While immunomagnetic cell separation has been utilized in all these areas, it has been hampered by the relatively large size of the paramagnetic label as compared to the target cell and by the relatively crude devices used for separation. This study will have two primary goals: the development of a novel magnetic cell label, and the analysis and design of a continuous, magnetic cell sorter using this label. This new magnetic label consists of ferritin combined to monoclonal antibodies against a particular surface antigen on the target cell. Ferritin is an iron storage protein in mammals. The properties of ferritin labels will be characterized using a number of techniques including: nuclear magnetic resonance spectroscopy, atomic absorption spectroscopy, and transmission electron microscopy. A unique sandwich labeling technique combining fluorescent and magnetic label will be used in combination with flow cytometry to characterize the cell-label binding. Human peripheral lymphocytes and human breast carcinoma derived cells will be used in this study, in combination with monoclonal antibodies against the specific markers: CD3, CD4, CD8, CD16, CD34 and Ber-EP4. These markers identify cells important for the bone marrow transplantation therapy: T cell, helper/inducer, cytotoxic/suppressor, natural killer, stem and breast carcinoma cells, respectively. Due to the complexity involved, the analysis and design of a continuous, magnetic cell sorter will require the use of 3-Dimensional Particle Image Velocimetry (3-D PIV). 3-D PIV, developed at the Ohio State University, the proposed subcontract site of this study, will allow the determination of 3-D velocity vectors of labeled and unlabeled cells in high magnetic fields. The experimental analysis, combined with the relevant equations of motion, will provide information on the system at a micron scale. With this information an optimized cell sorter will be developed. In addition, it will be possible to design more advanced sorters, such as multistage system, in which very high purity or very high selectivity can be obtained in a relatively short time. The ultimate goal is to develop a continuous, magnetic cell sorter for large-scale separation for bone marrow transplantation therapies. Such a sorter will be applicable to rapid, large-scale T cell depletion, cancer cell purging and hematopoietic stem cell enrichment.