肿瘤细胞异质性介导的耐药已被公认为是肿瘤医治的绊脚石。因此,以高通量的方式在单细胞水平上阐明抗癌药物的细胞毒性对于开发精准治疗具有重要价值。然而,目前的技术在动态表征呈现给多种药物条件的数千个单细胞或细胞克隆的反应方面存在局限性。
我们开发了一个新的基于微流体的“SMART”平台,操作简单,能够产生大量的单细胞阵列和多重药物浓度,能够保持细胞存活,在微室中可保留和跟踪。这些功能是通过集成一个微流体室阵列(4320个单位)和一个六浓度梯度发生器(MAC)来实现的,这使得能够以高通量方式高效分析白血病药物对单细胞和细胞克隆的影响。
一个简单的程序产生6个芯片上的药物梯度来处理3000多个单细胞或单细胞衍生的克隆,因此可以有效和精确地分析细胞异质性。统计结果表明,伊马替尼(Ima)和白藜芦醇(Res)联合治疗单细胞或克隆比Ima或Res单药治疗更有效。显著降低的半数最大抑制浓度(IC50)表明。此外,与单细胞相比,单细胞来源的克隆在每种药物处理中显示更高的IC50。此外,从两名白血病患者分离的原代细胞在MAC上进行药物治疗后也发现具有明显的异质性。
这个基于微流体的“SMART”平台允许高通量的单细胞捕获和培养,动态药物梯度治疗和细胞反应监测,这代表了一种有效研究抗癌药物作用的新方法,应该有利于白血病和其他癌症的药物发现。
Tumor cell heterogeneity mediated drug resistance has been recognized as the stumbling block of cancer treatment. Elucidating the cytotoxicity of anticancer drugs at single-cell level in a high-throughput way is thus of great value for developing precision therapy. However, current techniques suffer from limitations in dynamically characterizing the responses of thousands of single cells or cell clones presented to multiple drug conditions.
We developed a new microfluidics-based \"SMART\" platform that is Simple to operate, able to generate a Massive single-cell array and Multiplex drug concentrations, capable of keeping cells Alive, Retainable and Trackable in the microchambers. These features are achieved by integrating a Microfluidic chamber Array (4320 units) and a six-Concentration gradient generator (MAC), which enables highly efficient analysis of leukemia drug effects on single cells and cell clones in a high-throughput way.
A simple procedure produces 6 on-chip drug gradients to treat more than 3000 single cells or single-cell derived clones and thus allows an efficient and precise analysis of cell heterogeneity. The statistic results reveal that Imatinib (Ima) and Resveratrol (Res) combination treatment on single cells or clones is much more efficient than Ima or Res single drug treatment, indicated by the markedly reduced half maximal inhibitory concentration (IC50). Additionally, single-cell derived clones demonstrate a higher IC50 in each drug treatment compared to single cells. Moreover, primary cells isolated from two leukemia patients are also found with apparent heterogeneity upon drug treatment on MAC.
This microfluidics-based \"SMART\" platform allows high-throughput single-cell capture and culture, dynamic drug-gradient treatment and cell response monitoring, which represents a new approach to efficiently investigate anticancer drug effects and should benefit drug discovery for leukemia and other cancers.