Cell Dispensing in Low-Volume Range with the Immediate Drop-on-Demand Technology (I-DOT)
Cell Dispensing in Low-Volume Range with the Immediate Drop-on-Demand Technology (I-DOT)
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DOI：
10.1177/2211068214562450
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发表时间：
2015-04
期刊：
影响因子：
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通讯作者：
L. Schober;Evy Büttner;Christopher Laske;Andrea Traube;Tobias Brode;Andrea Traube;T. Bauernhansl
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文献类型：
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作者：
L. Schober;Evy Büttner;Christopher Laske;Andrea Traube;Tobias Brode;Andrea Traube;T. Bauernhansl
细胞的处理和给药包括用于筛选和毒性测试的基于细胞的测定系统的微制造中最关键的步骤。因此，开发了即时按需滴加技术（I-DOT），以提供一种灵活的非接触式液体处理系统，能够在没有交叉污染风险的情况下将细胞和液体分配到纳升范围内的精确体积。液体通过短压缩空气脉冲从底部喷嘴内的源板分配，该脉冲通过快速释放阀提供到井中，从而超过喷嘴中的毛细压力。可以将限定体积的液滴直接点样到微孔板或其他细胞培养装置上。我们提出了一项研究的性能和生物学影响的这项技术，通过应用细胞系MCF-7，人成纤维细胞，和人间充质干细胞（hMSCs）。对于测试的所有细胞类型，分配后的活力与对照相当，并且在不存在凋亡细胞的情况下表现出相似的增殖率，并且hMSC的分化潜力未受损。即时按需滴注技术能够实现准确的细胞剂量，并为单细胞应用提供了有前途的潜力。
Handling and dosing of cells comprise the most critical step in the microfabrication of cell-based assay systems for screening and toxicity testing. Therefore, the immediate drop-on-demand technology (I-DOT) was developed to provide a flexible noncontact liquid handling system enabling dispensing of cells and liquid without the risk of cross-contamination down to a precise volume in the nanoliter range. Liquid is dispensed from a source plate within nozzles at the bottom by a short compressed air pulse that is given through a quick release valve into the well, thus exceeding the capillary pressure in the nozzle. Droplets of a defined volume can be spotted directly onto microplates or other cell culture devices. We present a study on the performance and biological impact of this technology by applying the cell line MCF-7, human fibroblasts, and human mesenchymal stem cells (hMSCs). For all cell types tested, viability after dispensing is comparable to the control and exhibits similar proliferation rates in the absence of apoptotic cells, and the differentiation potential of hMSCs is not impaired. The immediate drop-on-demand technology enables accurate cell dosage and offers promising potential for single-cell applications.