Dielectrophoretic enrichment of live chemo-resistant circulating-like pancreatic cancer cells from media of drug-treated adherent cultures of solid tumors.
Dielectrophoretic enrichment of live chemo-resistant circulating-like pancreatic cancer cells from media of drug-treated adherent cultures of solid tumors.
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介电泳富集来自实体瘤药物处理贴壁培养物的活的化学抗性循环样胰腺癌细胞。
DOI：
10.1039/d3lc00804e
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发表时间：
2024-01-30
期刊：
影响因子：
6.1
通讯作者：
中科院分区：
文献类型：
--
作者：
由于液体活检中循环肿瘤细胞（CTC）的数量较低，因此人们对从体外肿瘤培养物中富集替代性循环样间充质癌细胞亚群以用于分子谱分析和药物筛选非常感兴趣。释放到药物处理的贴壁癌细胞培养物的培养基中的活癌细胞表现出失巢凋亡抗性或远离其细胞外基质与营养源和废物汇的锚定非依赖性存活，这是转移的先决条件。从肿瘤培养物中富集这些细胞亚群可以潜在地用作循环样癌细胞的体外来源，与CTC相比具有更大的扩大潜力。然而，这些活的循环样癌细胞亚群在培养基中表现出与坏死和凋亡细胞的大小重叠，这使得选择性地富集它们，同时保持它们处于悬浮状态具有挑战性。我们提出了优化的流通高频（1 MHz）正介电电泳（pDEP）设备与连续的3D场的非均匀性，使富集的活的化疗耐药循环癌细胞亚群的转移性患者来源的胰腺肿瘤细胞的体外培养。该策略的核心是利用具有由监督机器学习设置的门的单细胞阻抗细胞术，以优化pDEP的频率，使得基于多个生物物理度量（包括膜生理学、细胞质电导率和细胞大小）选择活循环细胞，这是不可能使用仅基于细胞大小的确定性横向位移的。使用具有低水平的活循环细胞（<3%）的典型药物处理的样品，我们呈现了在20分钟内靶亚群的pDEP富集至约44%的水平，同时拒绝>90%的死细胞。这种利用单细胞阻抗细胞仪来指导介电电泳优化的策略对其他复杂的生物样品具有影响。提出了使用单细胞阻抗细胞术对癌细胞亚群的介电电泳分离进行优化，其中机器学习用于门控其阻抗度量。
Due to low numbers of circulating tumor cells (CTCs) in liquid biopsies, there is much interest in enrichment of alternative circulating-like mesenchymal cancer cell subpopulations from in vitro tumor cultures for utilization within molecular profiling and drug screening. Viable cancer cells that are released into the media of drug-treated adherent cancer cell cultures exhibit anoikis resistance or anchorage-independent survival away from their extracellular matrix with nutrient sources and waste sinks, which serves as a pre-requisite for metastasis. The enrichment of these cell subpopulations from tumor cultures can potentially serve as an in vitro source of circulating-like cancer cells with greater potential for scale-up in comparison with CTCs. However, these live circulating-like cancer cell subpopulations exhibit size overlaps with necrotic and apoptotic cells in the culture media, which makes it challenging to selectively enrich them, while maintaining them in their suspended state. We present optimization of a flowthrough high frequency (1 MHz) positive dielectrophoresis (pDEP) device with sequential 3D field non-uniformities that enables enrichment of the live chemo-resistant circulating cancer cell subpopulation from an in vitro culture of metastatic patient-derived pancreatic tumor cells. Central to this strategy is the utilization of single-cell impedance cytometry with gates set by supervised machine learning, to optimize the frequency for pDEP, so that live circulating cells are selected based on multiple biophysical metrics, including membrane physiology, cytoplasmic conductivity and cell size, which is not possible using deterministic lateral displacement that is solely based on cell size. Using typical drug-treated samples with low levels of live circulating cells (<3%), we present pDEP enrichment of the target subpopulation to ∼44% levels within 20 minutes, while rejecting >90% of dead cells. This strategy of utilizing single-cell impedance cytometry to guide the optimization of dielectrophoresis has implications for other complex biological samples. Optimization of dielectrophoretic separation of cancer cell subpopulations using single-cell impedance cytometry is presented, with machine learning used to gate their impedance metrics.
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影响因子：
254.7
作者：
Jemal, Ahmedin;Siegel, Rebecca;Thun, Michael J.
通讯作者：
Thun, Michael J.
影响因子：
5.8
作者：
Edd, Jon F.;Mishra, Avanish;Smith, Kyle C.;Kapur, Ravi;Maheswaran, Shyamala;Haber, Daniel A.;Toner, Mehmet
通讯作者：
Toner, Mehmet
影响因子：
4.6
作者：
Ip CK;Li SS;Tang MY;Sy SK;Ren Y;Shum HC;Wong AS
通讯作者：
Wong AS
影响因子：
4
作者：
Dillekas, Hanna;Rogers, Michael S.;Straume, Oddhjorn
通讯作者：
Straume, Oddhjorn
DOI：
10.1039/c2ib20171b
发表时间：
2012-10
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
Lu J;Barrios CA;Dickson AR;Nourse JL;Lee AP;Flanagan LA
通讯作者：
Flanagan LA