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Magnetic nanoparticle tagging and application of magnetophoresis to cellular therapy and imaging.

机译：磁性纳米颗粒标记以及磁泳在细胞治疗和成像中的应用。

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The development of the nanotechnology enables special tailoring the functional properties of nanoparticles for biomedical and diagnostic applications. In this dissertation, cell magnetic nanoparticle tagging and cell magnetophoresis are investigated for applications in the areas of cellular therapy and imaging. Enrichment of blood progenitor cells with concomitant depletion of unwanted cells improves patient recovery in autologous and allogeneic stem cell transplantations. In this study, a quadrupole magnetic flow sorter (QMS), a flow-through magnetophoretic device, was evaluated using two strategies for the enrichment of blood progenitors from the clinical leukapheresis product samples. In the positive selection, seven commercial CD34+ progenitor cell-labeling reagents were evaluated quantitatively for the best performance through analysis of magnetophoresis process using cell tracking velocimetry (CTV). The reagent that produced the highest magnetophoretic mobility was selected for positive selection experiments (n=22). The CD34+ cells were isolated with a purity of 60%-96%, a recovery of 18%-60% and an enrichment rate of 12-169. In the negative depletion, a tetrameric antibody cocktail (TAC) and magnetic colloid against non-progenitor leukocytes was evaluated and used for negative progenitor enrichment experiments (n=23). The progenitor cells were isolated with a purity of 30%-85%, a recovery of 49%-84% and an enrichment rate of 8-104. Negative cell retention, a critical issue for the QMS process scale-up, was also investigated using a full-factorial experimental design, followed by a regression analysis of the cell sorting process. The intracellular magnetic particle tagging has been recently proposed for in-vivo magnetic resonance imaging (MRI) tracking of magnetically labeled progenitor cells. In this dissertation, the magnetophoretic motion analysis is applied to quantitate the cellular particle uptake and intracellular iron content. The intracellular superparamagnetic iron oxide (SPIO) nanoparticle uptake was facilitated by transfection agents (TAs) in live cells. The uptake was measured quantitatively from magnetophoresis by CTV. Seven different TAs-SPIO complexes based on dendrimer, lipid and polyethylenimine compounds applied to mesenchymal and hematopoietic progenitor cells and cardiac fibroblasts were evaluated. The results showed significant labeling differences, important for reagent selection as an MRI intracellular contrast.
机译：纳米技术的发展使得能够为生物医学和诊断应用特别定制纳米颗粒的功能特性。本文研究了细胞磁性纳米颗粒标记和细胞磁泳在细胞治疗和成像领域的应用。在自体和同种异体干细胞移植中，血液祖细胞的富集伴随着有害细胞的消耗得以改善，从而提高了患者的康复率。在这项研究中，使用两种策略从临床白细胞分离术产品样本中富集血液祖细胞，对四极磁流分选器（QMS）（流通磁磁装置）进行了评估。在阳性选择中，通过使用细胞跟踪测速法（CTV）分析磁泳过程，对7种市售CD34 +祖细胞标记试剂的最佳性能进行了定量评估。选择产生最高磁致动性的试剂进行阳性选择实验（n = 22）。分离出CD34 +细胞，其纯度为60％-96％，回收率为18％-60％，富集率为12-169。在阴性消耗中，评估了针对非祖白细胞的四聚体抗体混合物（TAC）和磁性胶体，并将其用于阴性祖细胞富集实验（n = 23）。分离出祖细胞，纯度为30％-85％，回收率为49％-84％，富集率为8-104。负细胞保留是QMS扩大规模的关键问题，还使用全因子实验设计进行了研究，然后对细胞分选过程进行了回归分析。近来已经提出了细胞内磁性颗粒标记用于磁性标记的祖细胞的体内磁共振成像（MRI）跟踪。本文通过磁致动运动分析来定量细胞颗粒的摄取和细胞内铁的含量。活细胞中的转染剂（TAs）促进了细胞内超顺磁性氧化铁（SPIO）纳米颗粒的吸收。通过CTV从磁泳定量地测量摄取。评估了基于树状聚合物，脂质和聚乙烯亚胺化合物的7种不同的TAs-SPIO复合物，这些复合物应用于间充质和造血祖细胞和心脏成纤维细胞。结果显示出明显的标记差异，这对于作为MRI细胞内对比剂的试剂选择非常重要。

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