Abstract:
The precise and effective isolation of living circulating tumor cells (CTCs) from peripheral blood, followed by their real-time monitoring, is crucial for diagnosing cancer patients. In this study, a cell-imprinted double-network (DN) hydrogel modified with circular multi-DNA (CMD), coined the CMD-imprinted hydrogel with fixed cells as templates (CMD-CIDH), was developed. The hydrogel featured a customized surface for proficient capture of viable CTCs and in situ real-time fluorescent detection without subsequent release. The customized surface, constructed using polyacrylamide/chitosan DN hydrogel as the matrix on the cell template, had a dense network structure, thereby ensuring excellent stability and a low degradation rate. Optimal capture efficiencies, recorded at 93 ± 3% for MCF-7 cells and 90 ± 2% for Hela cells, were achieved by grafting the CMD and adjusting the nodule size on the customized surface. The capture efficiency remained significantly high at 67 ± 11% in simulated breast cancer patient experiments even at a minimal concentration of 5 cells mL-1. Furthermore, CMD grafted onto the surface produced a potent fluorescence signature, enabling in situ real-time fluorescent detection of the target cell\'s growth state even in complex environments. The customized surface is highly efficient for screening CTCs in peripheral blood and has promising potential for setting up the CTCs culture.
摘要:
从外周血中精确有效地分离活的循环肿瘤细胞(CTC),随后是他们的实时监控,对癌症患者的诊断至关重要。在这项研究中,用环状多DNA(CMD)修饰的细胞印迹双网络(DN)水凝胶,用固定细胞作为模板创造了CMD印迹水凝胶(CMD-CIDH),已开发。水凝胶具有定制的表面,可有效捕获活的CTC并进行原位实时荧光检测,而无需随后释放。定制的表面,以聚丙烯酰胺/壳聚糖DN水凝胶为基质在细胞模板上构建,具有密集的网络结构,从而确保优异的稳定性和低降解率。最佳捕获效率,MCF-7细胞记录为93±3%,Hela细胞记录为90±2%,通过移植CMD并调整定制表面上的结节尺寸来实现。在模拟的乳腺癌患者实验中,甚至在5个细胞mL-1的最小浓度下,捕获效率仍然显著高,为67±11%。此外,接枝到表面的CMD产生了有效的荧光特征,即使在复杂环境中,也能对靶细胞的生长状态进行原位实时荧光检测。定制的表面对于筛选外周血中的CTC是高效的,并且具有建立CTC培养物的有希望的潜力。