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J Cell Physiol. 2018 Sep;233(9):6327-6336. doi: 10.1002/jcp.26519. Epub 2018 Mar 25.

Analysis of tumoral spheres growing in a multichamber microfluidic device.

Author information

1
Instituto de Oncología Angel H. Roffo, Universidad de Buenos Aires, Buenos Aires, Argentina.
2
Fellow at Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
3
Universidad Tecnológica Nacional (UTN), Facultad Regional de Haedo, Paris, Buenos Aires, Argentina.
4
Millipore Sigma Corporation, Hayward, California.
5
Bio-MEMS and Microsystem Lab, Department of Electrical Engineering, University of South Florida, Tampa, Florida.
6
Instituto de Ingeniería Biomédica, Facultad de Ingeniería, Universidad de Buenos Aires, (UBA), Buenos Aires, Argentina.
7
Member at Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET), Buenos Aires, Argentina.

Abstract

Lab on a Chip (LOC) farming systems have emerged as a powerful tool for single cell studies combined with a non-adherent cell culture substrate and single cell capture chips for the study of single cell derived tumor spheres. Cancer is characterized by its cellular heterogeneity where only a small population of cancer stem cells (CSCs) are responsible for tumor metastases and recurrences. Thus, the in vitro strategy to the formation of a single cell-derived sphere is an attractive alternative to identify CSCs. In this study, we test the effectiveness of microdevices for analysis of heterogeneity within CSC populations and its interaction with different components of the extracellular matrix. CSC could be identify using specific markers related to its pluripotency and self-renewal characteristics such as the transcription factor Oct-4 or the surface protein CD44. The results confirm the usefulness of LOC as an effective method for quantification of CSC, through the formation of spheres under conditions of low adhesion or growing on components of the extracellular matrix. The device used is also a good alternative for evaluating the individual growth of each sphere and further identification of these CSC markers by immunofluorescence. In conclusion, LOC devices have not only the already known advantages, but they are also a promising tool since they use small amounts of reagents and are under specific culture parameters. LOC devices could be considered as a novel technology to be used as a complement or replacement of traditional studies on culture plates.

KEYWORDS:

CD44; Oct-4; cancer stem cell; lab on a chip; sphere formation assay; tumoral sphere

PMID:
29574936
DOI:
10.1002/jcp.26519

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