Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, USA.
Department of Internal Medicine, Division of Hematology, The Ohio State University, Columbus, OH, USA.
Biotechnol J. 2017 Sep;12(9). doi: 10.1002/biot.201600750. Epub 2017 Aug 14.
Immunomagnetic separation is used to isolate circulating endothelial cells (ECs) and endothelial progenitor cells (EPCs) for diagnostics and tissue engineering. However, potentially detrimental changes in cell properties have been observed post-separation. Here, the effect of mechanical force, which is naturally applied during immunomagnetic separation, on proliferation of human umbilical vein endothelial cells (HUVEC), kinase insert domain-positive receptor (KDR) cells, and peripheral blood mononuclear cells (PBMCs). Cells are exposed to CD31 or Vascular Endothelial Growth Factor Receptor-2 (VEGFR2) targeted MACSi beads at varying bead to cell ratios and compared to free antibody and unconjugated beads. A vertical magnetic gradient is applied to static 2D cultures, and a magnetic cell sorter is used to analyze cells in dynamic flow. No significant difference in EC proliferation is observed for controls or VEGFR2-targeting beads, whereas CD31-conjugated beads increase proliferation in a dose dependent manner in static 2-D cultures. This effect occurs in the absence of magnetic field, but is more pronounced with magnetic force. After flow sorting, similar increases in proliferation are seen for CD31 targeting beads. Thus, the effects of targeting antibody and magnetic force applied should be considered when designing immunomagnetic separation protocols for ECs.
免疫磁珠分离技术用于分离循环内皮细胞 (ECs) 和内皮祖细胞 (EPCs)，用于诊断和组织工程。然而，在分离后观察到细胞特性的潜在有害变化。在这里，研究了免疫磁珠分离过程中自然施加的机械力对人脐静脉内皮细胞 (HUVEC)、激酶插入结构域阳性受体 (KDR) 细胞和外周血单核细胞 (PBMC) 增殖的影响。将细胞暴露于针对 CD31 或血管内皮生长因子受体-2 (VEGFR2) 的 MACSi 珠，以不同的珠与细胞比，并与游离抗体和未结合的珠进行比较。将垂直磁场施加于静态 2D 培养物上，并使用磁细胞分选器分析动态流动中的细胞。对于对照或针对 VEGFR2 的珠，未观察到 EC 增殖的显著差异，而 CD31 缀合珠在静态 2-D 培养物中以剂量依赖性方式增加增殖。这种效应发生在没有磁场的情况下，但在磁场力的作用下更为明显。在流式分选后，对于 CD31 靶向珠也观察到类似的增殖增加。因此，在设计用于 ECs 的免疫磁珠分离方案时，应考虑靶向抗体和施加的磁力的影响。
