1. Systems Biology for Improved In vitro Expansion of Cancer Stem Cells

Lab Objective

2. Glioblastoma Stem Cell Media Optimization

Cost of GSC media

Glioblastoma stem cells (GSCs) are typically cultured in vitro in serum-free Neurobasal-A media (NBE) containing a variety of supplements, the most expensive of which is basic fibroblast growth factor (bFGF). Growing cells in this manner is difficult and expensive, so culture methods need to be improved to make greater numbers of GSCs accessible in the future. This project explores the degradation rate of bFGF in cell culture, as well as its effect on stemness to confirm its necessity in culture and to determine the optimal amount for GSC in vitro growth. In addition, we are characterizing the relationship between culture pH and bFGF and analyzing how these factors work together to impact GSC growth. Through this research, we hope to develop an optimal media composition for GSCs that will increase the number of cells available for future research applications.

3. The Effects of Y-27632 on the In Vitro Expansion of Glioblastoma Stem Cells

Y-27632

This project explores the effects of the Rho-associated protein kinase inhibiting pyridine derivative compound Y-27632 on U87-MG glioblastoma tumorspheres. U87-MG cells form tumorspheres when grown in serum-free media that are enriched for glioblastoma stem cells (GSCs). Y-27632 has been implicated in suppressing apoptosis and cell-to-cell adhesion. It is hypothesized that Y-27632 would prevent apoptosis, increase stem cell populations, and negatively affect the cells’ ability to form tumorspheres. We are observing that Y-27632 increases GSC populations and have an overall positive effect on U87-MG tumorsphere formation and is poised to have promising application for cancer stem cell culture.

4. Characterization of the Extracellular Matrix of Glioblastoma Tumorspheres

GBM Tumor Sphere

The goal of this project is to characterize the extracellular matrix (ECM) of glioblastoma cells grown in multicellular spheroids or tumorspheres. Cancer cells are thought to excrete an extremely dense ECM which prevents even small molecules such as chemotherapeutics from penetrating into the cells. We are interested in exploring the composition of the ECM secreted by cells grown in vitro. This will mainly be accomplished through immunocytochemical staining with a fluorescence microscope. We also plan examine cells grown in different conditions, i.e. tumorspheres and hydrogels of different stiffnesses to understand how the ECM changes in different conditions, how these different conditions affect the glioblastoma stem cell population, and how the conditions affect the cells’ response to chemotherapeutic drugs. We seek to determine through this research which in vitro growth condition best models an in vivo tumor.

5. Fluid Shear Stress Metastasis Model for the Study of Cancer Stem Cells and Circulating Tumor Cells

Metastasis Model

While cancer is the second leading cause of death in the U.S., most cancer deaths are due to metastasis where the cancer spreads from the initial tumor location to other organs in the body by the vascular system. This project focuses on simulating the fluid shear stress experienced by cancer cells in the human vascular system during metastasis. We developed a cost-effective microfluidic system and are testing how cancer stem cell and circulating tumor cell (CTC) traits are impacted under physiological fluid shear stress. By looking at these traits of cancer cells, we hope to gain an understanding of the cancer cell during metastasis.

Last updated: 12-5-2014
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