Label-free microfluidic enrichment of cancer cells from noncancer cells in ascites fluid

 描述(由申请人提供)：对从积液和腹水中收集的癌细胞的检查是病理学家识别或确定癌症进展的重要步骤，也是癌症科学家准备执行分析技术的重要步骤。然而，由于存在许多非肿瘤细胞，包括间皮细胞、巨噬细胞和淋巴细胞，这项检查很复杂。可以利用诸如离心法之类的标准制备技术来增加样本中的总细胞密度，但进一步纯化收集的样本以高度浓缩癌细胞是麻烦和昂贵的，通常需要费力的处理和昂贵的标记试剂，如抗体。因此，从腹水中提纯癌细胞在临床上很少进行，在研究实验室中也很少见。这项研究提案的目的是测试标签 自由微流控细胞分选技术，利用细胞硬度和细胞大小的差异，从腹水中存在的非癌细胞中浓缩癌细胞。这项技术利用了观察到的事实，即癌细胞经常经历生物物理和形态变化，以区别于其他类型的非癌细胞。这项技术由带有周期性收缩的微通道组成，这些微通道旨在引导软细胞沿着不同于STIF细胞的路径。我们将研究如何利用通道几何形状和其他系统参数的变化，通过固有的生物物理差异来刺激癌细胞分离。一种高通量、无标记的方法从非癌细胞中提纯癌细胞，将大大提高癌症科学和临床诊断的准确性和效率。该项目的成功完成将创造一种新的浓缩方法，以促进检测复杂液体中极低水平的病变细胞，或提高包括积液和腹水在内的不同混合物中癌细胞亚群的纯度。

 DESCRIPTION (provided by applicant): The examination of cancer cells collected from effusions and ascites is an important step to identify or determine the progress of cancer by pathologists or in preparation of performing analytical techniques by cancer scientists. However this examination is complicated due to the presence of many noncancerous cells, including mesothelial, macrophage, and lymphocyte cells. Standard preparation techniques such as centrifugation can be utilized to increase the density of total cells in the sample, but further purification of the collected sample to highly enrich the cancer cells is troublesome and expensive, typically requiring laborious processing and expensive labeling reagents such as antibodies. As such, enriching cancer cells from ascites is rarely performed in the clinic and infrequently in the research laboratory. The objective of this research proposal is to test a label free microfluidic cell sorting technology that utilizes differences in cell stiffness and cell sizeto enrich cancer cells from noncancerous cells present in ascites fluid. The technology utilizes the observed fact that the cancer cells frequently undergo biophysical and morphological changes to be distinct from other noncancerous cell types. The technology consists of microchannels with periodic constrictions that are designed to direct soft cells along a different path than stif cells. We will examine how variations in channel geometry and other system parameters can be harnessed to provoke cancer cell segregation through inherent biophysical differences. A high throughput and label-free method to purify cancer cells from noncancerous cells will greatly benefit the accuracy and efficiency of both cancer science and clinical diagnoses. Successful completion of this project will create a new enrichment method to facilitate the detection of very low levels of diseased cells in complex fluids or to enhance the purity of cancer cell subpopulations within heterogeneous mixtures, including effusions and ascites fluid.