Ex vivo expansion of circulating lung tumor cells based on one-step microfluidics-based immunomagnetic isolation

Wang, Zhihua; Wu, Wenjun; Wang, Zhuo; Tang, Ying; Deng, Yuliang; Xu, Ling; Tian, Jie; Shi, Qi

doi:10.1039/c5an02554k

Cited by 34 publications

(39 citation statements)

References 27 publications

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“…After a wash (20 mL/h, 2 min) with HBSS to remove nonspecifically bound blood cells, magnets were retracted and tumor cells were released and collected in a ∼200 μL of solution. 16,17 …”

Section: Methodsmentioning

confidence: 99%

Single-Cell, Multiplexed Protein Detection of Rare Tumor Cells Based on a Beads-on-Barcode Antibody Microarray

Liu¹,

Wang²,

Deng³

et al. 2016

Anal. Chem.

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Circulating tumor cells (CTCs) shed from tumor sites and represent the molecular characteristics of the tumor. Besides genetic and transcriptional characterization, it is important to profile a panel of proteins with single-cell precision for resolving CTCs’ phenotype, organ-of-origin, and drug targets. We describe a new technology that enables profiling multiple protein markers of extraordinarily rare tumor cells at the single- cell level. This technology integrates a microchip consisting of 15000 60 pL-sized microwells and a novel beads-on-barcode antibody microarray (BOBarray). The BOBarray allows for multiplexed protein detection by assigning two independent identifiers (bead size and fluorescent color) of the beads to each protein. Four bead sizes (1.75, 3, 4.5, and 6 μm) and three colors (blue, green, and yellow) are utilized to encode up to 12 different proteins. The miniaturized BOBarray can fit an array of 60 pL- sized microwells that isolate single cells for cell lysis and the subsequent detection of protein markers. An enclosed 60 pL-sized microchamber defines a high concentration of proteins released from lysed single cells, leading to single-cell resolution of protein detection. The protein markers assayed in this study include organ-specific markers and drug targets that help to characterize the organ-of-origin and drug targets of isolated rare tumor cells from blood samples. This new approach enables handling a very small number of cells and achieves single-cell, multiplexed protein detection without loss of rare but clinically important tumor cells.

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“…After a wash (20 mL/h, 2 min) with HBSS to remove nonspecifically bound blood cells, magnets were retracted and tumor cells were released and collected in a ∼200 μL of solution. 16,17 …”

Section: Methodsmentioning

confidence: 99%

Single-Cell, Multiplexed Protein Detection of Rare Tumor Cells Based on a Beads-on-Barcode Antibody Microarray

Liu¹,

Wang²,

Deng³

et al. 2016

Anal. Chem.

Self Cite

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“…One way to enable the molecular and functional analysis of the few CTCs that are present in circulation is to expand the CTCs ex-vivo . This has shown promise in breast, prostate and lung cancer where short term CTC culture has been performed [ 4 , 8 , 9 ]. Until recently, CTC expansion ex-vivo was the bottleneck in the field.…”

Section: Introductionmentioning

confidence: 99%

“…Until recently, CTC expansion ex-vivo was the bottleneck in the field. The need for new isolation technologies that preserve the CTCs integrity in order to perform functional analysis has been a recent emphasis [ 9 , 10 ]. Viable CTCs grown in culture can potentially be used for therapeutic screening and used to guide patient therapy.…”

Section: Introductionmentioning

confidence: 99%

Short term ex-vivo expansion of circulating head and neck tumour cells

et al. 2016

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Minimally invasive techniques are required for the identification of head and neck cancer (HNC) patients who are at an increased risk of metastasis, or are not responding to therapy. An approach utilised in other solid cancers is the identification and enumeration of circulating tumour cells (CTCs) in the peripheral blood of patients. Low numbers of CTCs has been a limiting factor in the HNC field to date. Here we present a methodology to expand HNC patient derived CTCs ex-vivo. As a proof of principle study, 25 advanced stage HNC patient bloods were enriched for circulating tumour cells through negative selection and cultured in 2D and 3D culture environments under hypoxic conditions (2% O2, 5% CO2). CTCs were detected in 14/25 (56%) of patients (ranging from 1–15 CTCs/5 mL blood). Short term CTC cultures were successfully generated in 7/25 advanced stage HNC patients (5/7 of these cultures were from HPV+ patients). Blood samples from which CTC culture was successful had higher CTC counts (p = 0.0002), and were predominantly from HPV+ patients (p = 0.007). This is, to our knowledge, the first pilot study to culture HNC CTCs ex-vivo. Further studies are warranted to determine the use of short term expansion in HNC and the role of HPV in promoting culture success.

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“…Under an external magnetic field, leukocytes labelled with anti-human CD45 antibodies-coated magnetic beads were removed, and target cells left behind were effectively enriched. Wang et al [ 161 ] reported the use of a herringbone microstructure in a microfluidic MBAT chip. The introduction of a herringbone microstructure produces two favorable effects: (i) the herringbone microstructure induces vortices which bring CTCs close to herringbone grooves.…”

Section: Other Microfluidic Concentration Methodsmentioning

confidence: 99%

Microfluidic Techniques for Analytes Concentration

Zhao

Yang

2017

Micromachines

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Microfluidics has been undergoing fast development in the past two decades due to its promising applications in biotechnology, medicine, and chemistry. Towards these applications, enhancing concentration sensitivity and detection resolution are indispensable to meet the detection limits because of the dilute sample concentrations, ultra-small sample volumes and short detection lengths in microfluidic devices. A variety of microfluidic techniques for concentrating analytes have been developed. This article presents an overview of analyte concentration techniques in microfluidics. We focus on discussing the physical mechanism of each concentration technique with its representative advancements and applications. Finally, the article is concluded by highlighting and discussing advantages and disadvantages of the reviewed techniques.

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Ex vivo expansion of circulating lung tumor cells based on one-step microfluidics-based immunomagnetic isolation

Cited by 34 publications

References 27 publications

Single-Cell, Multiplexed Protein Detection of Rare Tumor Cells Based on a Beads-on-Barcode Antibody Microarray

Single-Cell, Multiplexed Protein Detection of Rare Tumor Cells Based on a Beads-on-Barcode Antibody Microarray

Short term ex-vivo expansion of circulating head and neck tumour cells

Microfluidic Techniques for Analytes Concentration

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