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Removal of Human Leukemic Cells from Peripheral Blood Mononuclear Cells by Cell Recognition Chromatography with Size Matched Particle Imprints
ACS Applied Bio Materials ( IF 4.6 ) Pub Date : 2020-01-17 , DOI: 10.1021/acsabm.9b00770 Rosie Chester 1 , Anupam A K Das 1 , Jevan Medlock 1 , Dieter Nees 2 , David J Allsup 3 , Leigh A Madden 4 , Vesselin N Paunov 1
ACS Applied Bio Materials ( IF 4.6 ) Pub Date : 2020-01-17 , DOI: 10.1021/acsabm.9b00770 Rosie Chester 1 , Anupam A K Das 1 , Jevan Medlock 1 , Dieter Nees 2 , David J Allsup 3 , Leigh A Madden 4 , Vesselin N Paunov 1
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We report a cell recognition chromatography approach for blood cancer cell separation from healthy peripheral blood mononuclear cells (PBMCs) based on size-matched functionalized particle imprints. Negative imprints were prepared from layers of 15 μm polymeric microbeads closely matching the size of cultured human leukemic cells (HL60). We replicated these imprints on a large scale with UV curable polyurethane resin using nanoimprinting lithography. The imprints were functionalized with branched polyethylene imine (bPEI) and passivated by Poloxamer 407 to promote a weak attraction toward cells. When a matching cell fits into an imprint cavity, its contact area with the imprint is maximized, which amplifies the attraction and the binding selectivity. We tested these imprints specificity for depleting myeloblasts from a mixture with healthy human PBMCs in a cell recognition chromatography setup hosting the imprint. The mixture of fixed HL60/PBMCs ratio was circulated over the imprint and at each step the selectivity toward HL60 was assessed by flow cytometry. The role of the imprint length, flow rate, channel depth, and the bPEI coating concentration were examined. The results show that HL60 cells, closely matching the imprint cavities, get trapped on the imprint, while the smaller PBMCs are carried away by the drag force of the flow. Lower flow rates, longer imprints, and interim channel depth favor HL60 specific retention. The bPEI concentration higher than 1 wt % on the imprint made it less selective toward the HL60 because of indiscriminate attraction with all cells. Particle imprint based cell recognition chromatography was able to achieve selective myeloblast depletion from initial 11.7% HL60 (88.3% PBMC) to less than 1.3% HL60 for 3 h of circulation. The cell recognition chromatography with size-matched microbead imprints can be employed as an efficient cell separation technique and potentially lead to alternative therapies for myeloblasts removal from peripheral blood of patients with acute myeloid leukemia.
中文翻译：
通过具有大小匹配颗粒印记的细胞识别色谱法从外周血单个核细胞中去除人白血病细胞
我们报告了一种基于尺寸匹配的功能化粒子印记的细胞识别色谱方法，用于从健康的外周血单核细胞 (PBMC) 中分离血癌细胞。负印记由与培养的人类白血病细胞 (HL60) 大小密切匹配的 15 μm 聚合物微珠层制备。我们使用纳米压印光刻技术用紫外线固化聚氨酯树脂大规模复制了这些印记。印迹用支化聚乙烯亚胺 (bPEI) 官能化，并用泊洛沙姆 407 钝化以促进对细胞的弱吸引力。当匹配的细胞装入印记腔时，其与印记的接触面积最大化，从而放大了吸引力和结合选择性。我们测试了这些印记特异性，用于在承载印记的细胞识别色谱装置中从与健康人类 PBMC 的混合物中消耗成髓细胞。固定 HL60/PBMCs 比例的混合物在印记上循环，并且在每一步通过流式细胞术评估对 HL60 的选择性。检查了压印长度、流速、通道深度和 bPEI 涂层浓度的作用。结果表明，与印迹腔紧密匹配的 HL60 细胞被困在印迹上，而较小的 PBMC 则被流动的阻力带走。较低的流速、较长的印迹和临时通道深度有利于 HL60 的特定保留。印记上高于 1 wt% 的 bPEI 浓度使其对 HL60 的选择性降低，因为它与所有细胞不分青红皂白地吸引。基于粒子印记的细胞识别色谱能够在循环 3 小时内实现从最初的 11.7% HL60 (88.3% PBMC) 到小于 1.3% HL60 的选择性成髓细胞消耗。具有大小匹配的微珠印记的细胞识别色谱可用作有效的细胞分离技术，并可能导致从急性髓性白血病患者的外周血中去除成髓细胞的替代疗法。
更新日期：2020-01-17
中文翻译：
通过具有大小匹配颗粒印记的细胞识别色谱法从外周血单个核细胞中去除人白血病细胞
我们报告了一种基于尺寸匹配的功能化粒子印记的细胞识别色谱方法，用于从健康的外周血单核细胞 (PBMC) 中分离血癌细胞。负印记由与培养的人类白血病细胞 (HL60) 大小密切匹配的 15 μm 聚合物微珠层制备。我们使用纳米压印光刻技术用紫外线固化聚氨酯树脂大规模复制了这些印记。印迹用支化聚乙烯亚胺 (bPEI) 官能化，并用泊洛沙姆 407 钝化以促进对细胞的弱吸引力。当匹配的细胞装入印记腔时，其与印记的接触面积最大化，从而放大了吸引力和结合选择性。我们测试了这些印记特异性，用于在承载印记的细胞识别色谱装置中从与健康人类 PBMC 的混合物中消耗成髓细胞。固定 HL60/PBMCs 比例的混合物在印记上循环，并且在每一步通过流式细胞术评估对 HL60 的选择性。检查了压印长度、流速、通道深度和 bPEI 涂层浓度的作用。结果表明，与印迹腔紧密匹配的 HL60 细胞被困在印迹上，而较小的 PBMC 则被流动的阻力带走。较低的流速、较长的印迹和临时通道深度有利于 HL60 的特定保留。印记上高于 1 wt% 的 bPEI 浓度使其对 HL60 的选择性降低，因为它与所有细胞不分青红皂白地吸引。基于粒子印记的细胞识别色谱能够在循环 3 小时内实现从最初的 11.7% HL60 (88.3% PBMC) 到小于 1.3% HL60 的选择性成髓细胞消耗。具有大小匹配的微珠印记的细胞识别色谱可用作有效的细胞分离技术，并可能导致从急性髓性白血病患者的外周血中去除成髓细胞的替代疗法。
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