Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Chen, Yu-Chih, Ingram, Patrick N., Fouladdel, Shamileh, McDermott, Sean P., Azizi, Ebrahim, Wicha, Max S., Yoon, Euisik
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904376/
https://www.ncbi.nlm.nih.gov/pubmed/27292795
http://dx.doi.org/10.1038/srep27301
_version_ 1782437138300665856
author Chen, Yu-Chih
Ingram, Patrick N.
Fouladdel, Shamileh
McDermott, Sean P.
Azizi, Ebrahim
Wicha, Max S.
Yoon, Euisik
author_facet Chen, Yu-Chih
Ingram, Patrick N.
Fouladdel, Shamileh
McDermott, Sean P.
Azizi, Ebrahim
Wicha, Max S.
Yoon, Euisik
author_sort Chen, Yu-Chih
collection PubMed
description 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.
format Online
Article
Text
id pubmed-4904376
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-49043762016-06-14 High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis Chen, Yu-Chih Ingram, Patrick N. Fouladdel, Shamileh McDermott, Sean P. Azizi, Ebrahim Wicha, Max S. Yoon, Euisik Sci Rep Article 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. Nature Publishing Group 2016-06-13 /pmc/articles/PMC4904376/ /pubmed/27292795 http://dx.doi.org/10.1038/srep27301 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Chen, Yu-Chih
Ingram, Patrick N.
Fouladdel, Shamileh
McDermott, Sean P.
Azizi, Ebrahim
Wicha, Max S.
Yoon, Euisik
High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis
title High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis
title_full High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis
title_fullStr High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis
title_full_unstemmed High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis
title_short High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis
title_sort high-throughput single-cell derived sphere formation for cancer stem-like cell identification and analysis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904376/
https://www.ncbi.nlm.nih.gov/pubmed/27292795
http://dx.doi.org/10.1038/srep27301
work_keys_str_mv AT chenyuchih highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis
AT ingrampatrickn highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis
AT fouladdelshamileh highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis
AT mcdermottseanp highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis
AT aziziebrahim highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis
AT wichamaxs highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis
AT yooneuisik highthroughputsinglecellderivedsphereformationforcancerstemlikecellidentificationandanalysis