Microfluidic Device Development for Analyzing Single Glioblastoma Cells
- Author(s): Lin, Jung-ming G.
- Advisor(s): Kumar, Sanjay
- et al.
Glioblastoma remains a deadly disease due to the diffuse infiltration of single tumor
cells into the surrounding tissue. Even with the current treatment regimens of surgery,
radiation, and chemotherapy, the median survival time is approximately one year. Like
many solid tumors, glioblastoma is extremely heterogeneous with respect to multiple
phenotypes such as invasive capacity, therapeutic resistance, and tumorigenicity. This
heterogeneity complicates our understanding of glioblastoma and consequently, our
ability to treat this disease. Unfortunately, standard population-based assays can mask
the properties of rare subpopulations within a tumor and therefore, obscure our
understanding of these subpopulations. As a result, without tools that allow for single
cell analysis, we are unable to interrogate how different subpopulation phenotypes may
serve as indicators of glioblastoma tumor growth and progression.
In this dissertation, we sought to develop and optimize new microfluidic tools to analyze
single glioblastoma cells for a range of phenotypes: viscoelasticity, motility, and
invasion. First, we developed a cross-slot based platform and a corresponding
analytical model that enables the determination of cellular viscoelastic properties
(stiffness and fluidity) in a high-throughput manner. Using this platform, we quantified
the viscoelastic properties of 3T3 fibroblasts and glioblastoma tumor initiating cells
(TICs) and observed the expected changes in the cellular elastic modulus in response
to agents that soften or stiffen the cytoskeleton. Second, we developed a microfluidic
device and workflows that integrates measurements of invasive motility and targeted
protein expression with single cell resolution, which we named SCAMPR (Single Cell
Analysis of Motility and Proteotype). Using this platform, we identified two proteins,
Nestin and EphA2, which positively correlates with TIC invasive motility.
In summary, this dissertation focuses on the development of microfluidic platforms for
single cell analysis. Our developed platforms provide a method in which to interrogate
single cells in a high throughput manner and to identify novel relationships between
varied cellular phenotypes. These insights are crucial for the identification of both rare
subpopulations within a given tumor and patient-specific protein markers that describe