High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis

Considerable evidence suggests that many malignancies are driven by a cellular compartment that displays stem cell properties. Cancer stem-like cells (CSCs) can be identified by expression of cell surface markers or enzymatic activity, but these methods are limited by phenotypic heterogeneity and pl...

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Bibliographic Details
Published in:Scientific reports 2016-06, Vol.6 (1), p.27301-27301, Article 27301
Main Authors: Chen, Yu-Chih, Ingram, Patrick N, Fouladdel, Shamileh, McDermott, Sean P, Azizi, Ebrahim, Wicha, Max S, Yoon, Euisik
Format: Article
Language:English
Subjects:
Animals
Biomarkers, Tumor - genetics
Biomarkers, Tumor - metabolism
Breast cancer
Cell Line, Tumor
Female
Humans
Hydrogels - chemistry
Lab-On-A-Chip Devices
MCF-7 Cells
Mice
Mice, Inbred NOD
Mice, SCID
Microfluidics - instrumentation
Microfluidics - methods
Microscopy, Electron, Scanning
Neoplastic Stem Cells - cytology
Neoplastic Stem Cells - metabolism
Polyhydroxyethyl Methacrylate - chemistry
Receptors, Notch - genetics
Receptors, Notch - metabolism
Single-Cell Analysis - methods
Spheres
Spheroids, Cellular - cytology
Spheroids, Cellular - metabolism
Stem cells
Transplantation, Heterologous
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Summary:Considerable evidence suggests that many malignancies are driven by a cellular compartment that displays stem cell properties. Cancer stem-like cells (CSCs) can be identified by expression of cell surface markers or enzymatic activity, but these methods are limited by phenotypic heterogeneity and plasticity of CSCs. An alternative phenotypic methodology based on in-vitro sphere formation has been developed, but it is typically labor-intensive and low-throughput. In this work, we present a 1,024-microchamber microfluidic platform for single-cell derived sphere formation. Utilizing a hydrodynamic capturing scheme, more than 70% of the microchambers capture only one cell, allowing for monitoring of sphere formation from heterogeneous cancer cell populations for identification of CSCs. Single-cell derived spheres can be retrieved and dissociated for single-cell analysis using a custom 96-gene panel to probe heterogeneity within the clonal CSC spheres. This microfluidic platform provides reliable and high-throughput sphere formation for CSC identification and downstream clonal analysis.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep27301