Department of Bioengineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Gunma Prefectural Textile Industrial Laboratory, 5-46-1 Aioicho, Kiryu, Gunma 376-0011, Japan.
Acta Biomater. 2018 Feb;67:32-41. doi: 10.1016/j.actbio.2017.11.055. Epub 2017 Dec 6.
Circulating tumor cells (CTCs) are tumor cells present in the blood. CTCs have attracted much attention as a new tumor marker, because their analysis provides useful information for monitoring cancer progress. In this study, we developed cell-capture and release methods using three-dimensional (3D) microfiber fabrics without damaging the cells. Using functional peptides containing sequences from a polystyrene-binding site and a cleavable site for collagenase type IV, immobilized antibodies on the peptides were able to specifically capture MCF-7 cells in a few minutes and release the captured cells from 3D microfiber fabrics incorporating a vacuum system. The efficiency of cell capture was around 80% and that of the cell release was over 90%. The released cells proliferated normally in culture medium, suggesting that our system will be applicable for the culture and analysis of CTCs.
In this paper, we report cell-capture and release methods using enzyme-cleavable peptides immobilized on microfiber fabrics which has microporous polymeric three-dimensional structures. Detachment and collection of the selectively captured cancer cells are required for ex vivo culture and their further analysis, whereas the cell detachment methods developed so far might cause cell damage, even if cell viability is high enough. Therefore, specific attachment and gentle detachment from the device are required for the accurate analysis of cells. In this study, for capture and release of cancer cells we designed the peptide cleavable by collagenase type IV, which has no target molecule in cells. Our system will be useful for further CTC analysis and might lead to more accurate cancer diagnosis.
循环肿瘤细胞(CTCs)是存在于血液中的肿瘤细胞。由于其分析为监测癌症进展提供了有用的信息,因此作为一种新的肿瘤标志物,CTCs 引起了广泛关注。在这项研究中,我们开发了使用三维(3D)微纤维织物的细胞捕获和释放方法,而不会对细胞造成损伤。使用含有聚苯乙烯结合位点和胶原酶 IV 可切割位点序列的功能肽,固定在肽上的抗体能够在几分钟内特异性捕获 MCF-7 细胞,并通过结合真空系统的 3D 微纤维织物从捕获的细胞中释放出来。细胞捕获效率约为 80%,细胞释放效率超过 90%。释放的细胞在培养基中正常增殖,表明我们的系统将适用于 CTC 的培养和分析。
在本文中,我们报告了使用固定在微纤维织物上的酶可切割肽的细胞捕获和释放方法,微纤维织物具有微孔聚合物三维结构。为了进行体外培养和进一步分析,需要分离和收集选择性捕获的癌细胞,而迄今为止开发的细胞分离方法可能会导致细胞损伤,即使细胞活力足够高。因此,需要对设备进行特异性附着和温和分离,以便对细胞进行准确分析。在这项研究中,我们设计了可被胶原酶 IV 切割的肽,用于捕获和释放癌细胞,该肽在细胞中没有靶分子。我们的系统将有助于进一步的 CTC 分析,并可能导致更准确的癌症诊断。