Abstract
Integration of microfabricated, single-cell resolution and traditional, population-level biological assays will be the future of modern techniques in biology that will enroll in the evolution of biology into a precision scientific discipline. In this study, we developed a microfabricated cell culture platform to investigate the indirect influence of macrophages on glioma cell behavior. We quantified proliferation, morphology, motility, migration, and deformation properties of glioma cells at single-cell level and compared these results with population-level data. Our results showed that glioma cells obtained slightly slower proliferation, higher motility, and extremely significant deformation capability when cultured with 50% regular growth medium and 50% macrophage-depleted medium. When the expression levels of E-cadherin and Vimentin proteins were measured, it was verified that observed mechanophenotypic alterations in glioma cells were not due to epithelium to mesenchymal transition. Our results were consistent with previously reported enormous heterogeneity of U87 glioma cell line. Herein, for the first time, we quantified the change of deformation indexes of U87 glioma cells using microfluidic devices for single-cells analysis.
Highlights
Glioblastoma or Glioblastoma multiforme (GBM), is the most devastating type of brain cancer with nonspecific signs and symptoms and very limited treatment strategies
We have investigated biomechanical properties of U87-MG (HTB-14 TM) glioma cells creating two glioma cell culturing conditions; i-U87 category: U87 glioma cells fed by the regular growth medium, and ii-U87 glioma conditional (U87-C) category: U87 glioma cells were cultured in 50% growth medium supplemented by the 50% macrophage-depleted medium
This study demonstrated a single-cell mechanophenotyping approach for U87 glioma cells while integrating microfluidic cell culture platforms with macroscale traditional assays
Summary
Glioblastoma or Glioblastoma multiforme (GBM), is the most devastating type of brain cancer with nonspecific signs and symptoms and very limited treatment strategies. It results in death in 15 months following diagnosis [1,2]. One of the main challenges for successful treatment of gliomas is its multiform structure [3,4]. Glioma multiforme is highly heterogeneous with different cell types and their complex interactions. It is an extremely dynamic, hierarchical microenvironment [5]. Increased aggressiveness and invasiveness of GBM requires adequate single-cell tools to avoid masking rare cells and to allow quantifying heterogeneity according to morphologic, phenotypic, and functional properties of glioma cells in a population
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