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基于传热方法并通过表面印迹聚合物进行细胞检测的细胞培养质量分析

Heat-transfer-method-based cell culture quality assay through cell detection by surface imprinted polymers.

作者信息

Eersels Kasper, van Grinsven Bart, Khorshid Mehran, Somers Veerle, Püttmann Christiane, Stein Christoph, Barth Stefan, Diliën Hanne, Bos Gerard M J, Germeraad Wilfred T V, Cleij Thomas J, Thoelen Ronald, De Ceuninck Ward, Wagner Patrick

机构信息

Hasselt University , Institute for Materials Research IMO, Wetenschapspark 1, Diepenbeek, Belgium.

出版信息

Langmuir. 2015 Feb 17;31(6):2043-50. doi: 10.1021/la5046173. Epub 2015 Feb 5.

Abstract

Previous work has indicated that surface imprinted polymers (SIPs) allow for highly specific cell detection through macromolecular cell imprints. The combination of SIPs with a heat-transfer-based read-out technique has led to the development of a selective, label-free, low-cost, and user-friendly cell detection assay. In this study, the breast cancer cell line ZR-75-1 is used to assess the potential of the platform for monitoring the quality of a cell culture in time. For this purpose, we show that the proposed methodology is able to discriminate between the original cell line (adherent growth, ZR-75-1a) and a descendant cell line (suspension growth, ZR-75-1s). Moreover, ZR-75-1a cells were cultured for a prolonged period of time and analyzed using the heat-transfer method (HTM) at regular time intervals. The results of these experiments demonstrate that the thermal resistance (Rth) signal decays after a certain number of cell culture passages. This can likely be attributed to a compromised quality of the cell culture due to cross-contamination with the ZR-75-1s cell line, a finding that was confirmed by classical STR DNA profiling. The cells do not express the same functional groups on their membrane, resulting in a weaker bond between cell and imprint, enabling cell removal by mechanical friction, provided by flushing the measuring chamber with buffer solution. These findings were further confirmed by HTM and illustrate that the biomimetic sensor platform can be used as an assay for monitoring the quality of cell cultures in time.

摘要

先前的研究表明,表面印迹聚合物(SIPs)可通过大分子细胞印迹实现高度特异性的细胞检测。SIPs与基于热传递的读出技术相结合,促成了一种选择性、无标记、低成本且用户友好的细胞检测方法的开发。在本研究中,乳腺癌细胞系ZR-75-1被用于评估该平台及时监测细胞培养质量的潜力。为此,我们证明了所提出的方法能够区分原始细胞系(贴壁生长,ZR-75-1a)和后代细胞系(悬浮生长,ZR-75-1s)。此外,将ZR-75-1a细胞长时间培养,并定期使用热传递方法(HTM)进行分析。这些实验结果表明,在细胞培养传代一定次数后,热阻(Rth)信号会衰减。这可能归因于与ZR-75-1s细胞系交叉污染导致的细胞培养质量受损,这一发现通过经典的STR DNA分析得到了证实。细胞在其膜上不表达相同的官能团,导致细胞与印迹之间的结合较弱,通过用缓冲溶液冲洗测量室提供的机械摩擦能够实现细胞去除。这些发现通过HTM得到了进一步证实,并表明仿生传感器平台可作为及时监测细胞培养质量的一种检测方法。

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