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Biotechniques. 2016 Jan 1;60(1):43-6. doi: 10.2144/000114372. eCollection 2016 Jan.

A simple, low-cost staining method for rapid-throughput analysis of tumor spheroids.

Author information

1
Robert H. Lurie Comprehensive Cancer Center and Division of Hematology-Oncology, Northwestern University Feinberg School of Medicine, Chicago, IL.
2
Department of Neurology, Robert H. Lurie Comprehensive Cancer Center, Northwestern Brain Tumor Institute, Northwestern University Feinberg School of Medicine, Chicago, IL.
3
Center for Advanced Microscopy and Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL.
4
Division of Hematology and Oncology, Ann & Robert H. Lurie Children's Hospital of Chicago, Chicago, IL.
5
Department of Medicine, Jesse Brown VA Medical Center, Chicago, IL.

Abstract

Tumor spheroids are becoming an important tool for the investigation of cancer stem cell (CSC) function in tumors; thus, low-cost and high-throughput methods for drug screening of tumor spheroids are needed. Using neurospheres as non-adherent three-dimensional (3-D) cultures, we developed a simple, low-cost acridine orange (AO)-based method that allows for rapid analysis of live neurospheres by fluorescence microscopy in a 96-well format. This assay measures the cross-section area of a spheroid, which corresponds to cell viability. Our novel method allows rapid screening of a panel of anti-proliferative drugs to assess inhibitory effects on the growth of cancer stem cells in 3-D cultures.

KEYWORDS:

acridine orange; cancer stem cell; glioblastoma; microscopy; neurospheres; tumor spheroids

PMID:
26757811
PMCID:
PMC4772731
DOI:
10.2144/000114372
[Indexed for MEDLINE]
Free PMC Article

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