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一种用于癌细胞分离和纯化的正负富集联合检测法。

A Combined Negative and Positive Enrichment Assay for Cancer Cells Isolation and Purification.

作者信息

Cheng Boran, Wang Shuyi, Chen Yuanyuan, Fang Yuan, Chen Fangfang, Wang Zhenmeng, Xiong Bin

机构信息

Department of Oncology, Zhongnan Hospital of Wuhan University, Hubei Key Laboratory of Tumor Biological Behaviors, Hubei Cancer Clinical Study Center, Wuhan, Hubei 430071, P. R. China.

Department of Oncology, Zhongnan Hospital of Wuhan University, Hubei Key Laboratory of Tumor Biological Behaviors, Hubei Cancer Clinical Study Center, Wuhan, Hubei 430071, P. R. China

出版信息

Technol Cancer Res Treat. 2016 Feb;15(1):69-76. doi: 10.7785/tcrt.2012.500447. Epub 2014 Nov 26.

Abstract

Cancer cells that detach from solid tumor and circulate in the peripheral blood (CTCs) have been considered as a new "biomarker" for the detection and characterization of cancers. However, isolating and detecting cancer cells from the cancer patient peripheral blood have been technically challenging, owing to the small sub-population of CTCs (a few to hundreds per milliliter). Here we demonstrate a simple and efficient cancer cells isolation and purification method. A biocompatible and surface roughness controllable TiO2 nanofilm was deposited onto a glass slide to achieve enhanced topographic interactions with nanoscale cellular surface components, again, anti-CD45 (a leukocyte common antigen) and anti-EpCAM (epithelial cell adhesion molecule) were then coated onto the surface of the nanofilm for advance depletion of white blood cells (WBCs) and specific isolation of CTCs, respectively. Comparing to the conventional positive enrichment technology, this method exhibited excellent biocompatibility and equally high capture efficiency. Moreover, the maximum number of background cells (WBCs) was removed, and viable and functional cancer cells were isolated with high purity. Utilizing the horizontally packed TiO2 nanofilm improved pure CTC-capture through combining cell-capture-agent and cancer cell-preferred nanoscale topography, which represented a new method capable of obtaining biologically functional CTCs for subsequent molecular analysis.

摘要

从实体瘤脱离并在外周血中循环的癌细胞(循环肿瘤细胞,CTCs)被视为癌症检测与特征分析的一种新型“生物标志物”。然而,从癌症患者外周血中分离和检测癌细胞在技术上颇具挑战,这是因为循环肿瘤细胞的亚群数量很少(每毫升只有几个到数百个)。在此,我们展示了一种简单高效的癌细胞分离与纯化方法。将具有生物相容性且表面粗糙度可控的二氧化钛纳米膜沉积在载玻片上,以增强与纳米级细胞表面成分的形貌相互作用,接着分别在纳米膜表面包被抗CD45(一种白细胞共同抗原)和抗EpCAM(上皮细胞粘附分子),用于预先去除白细胞(WBCs)和特异性分离循环肿瘤细胞。与传统的阳性富集技术相比,该方法具有出色的生物相容性和同样高的捕获效率。此外,去除了最大数量的背景细胞(白细胞),并以高纯度分离出有活力且功能正常的癌细胞。利用水平堆积的二氧化钛纳米膜,通过结合细胞捕获剂和癌细胞偏好的纳米级形貌,提高了对循环肿瘤细胞的纯捕获率,这代表了一种能够获取具有生物学功能的循环肿瘤细胞以用于后续分子分析的新方法。

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