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Antibody-Immobilized Column for Quick Cell Separation Based on Cell Rolling

机译：基于细胞滚动的固定化抗体的快速细胞分离柱

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摘要

Cell separation using methodological standards that ensure high purity is a very important step in cell transplantation for regenerative medicine and for stem cell research. A separation protocol using magnetic beads has been widely used for cell separation to isolate negative and positive cells. However, not only the surface marker pattern, e.g., negative or positive, but also the density of a cell depends on its developmental stage and differentiation ability. Rapid and label-free separation procedures based on surface marker density are the focus of our interest. In this study, we have successfully developed an antiCD34 antibody-immobilized cell-rolling column, that can separate cells depending on the CD34 density of the cell surfaces. Various conditions for the cell-rolling column were optimized including graft copolymerization, and adjustment of the column tilt angle, and medium flow rate. Using CD34-positive and -negative cell lines, the cell separation potential of the column was established. We observed a difference in the rolling velocities between CD34-positive and CD34-negative cells on antibody-immobilized microfluidic device. Cell separation was achieved by tilting the surface 20 degrees and the increasing medium flow. Surface marker characteristics of the isolated cells in each fraction were analyzed using a cell-sorting system, and it was found that populations containing high density of CD34 were eluted in the delayed fractions. These results demonstrate that cells with a given surface marker density can be continuously separated using the cell rolling column.
机译：使用可确保高纯度的方法学标准进行细胞分离，对于再生医学和干细胞研究而言，是细胞移植中非常重要的一步。使用磁珠的分离方案已被广泛用于细胞分离，以分离阴性和阳性细胞。然而，不仅表面标记图案，例如阴性或阳性，而且细胞的密度还取决于其发育阶段和分化能力。基于表面标记物密度的快速无标签分离程序是我们关注的焦点。在这项研究中，我们成功开发了固定有抗CD34抗体的细胞滚动柱，该柱可以根据细胞表面的CD34密度分离细胞。优化了细胞滚动柱的各种条件，包括接枝共聚，调节柱倾斜角和介质流速。使用CD34阳性和阴性细胞系，建立了柱的细胞分离潜能。我们观察到固定抗体的微流控设备上CD34阳性和CD34阴性细胞的滚动速度存在差异。通过将表面倾斜20度并增加培养基流量来实现细胞分离。使用细胞分选系统分析每个级分中分离出的细胞的表面标志物特征，发现在延迟级分中洗脱了含有高密度CD34的群体。这些结果表明，具有给定表面标记物密度的细胞可以使用细胞滚动柱连续分离。

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