Honors College Thesis
 

Formation and Migration of Breast Cancer Spheroids

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  • Invasive breast cancer affects 1 in 8 women during their lifetime. Current cancer research uses 2-D cell culturing, which does not accurately represent tumor behavior. In comparison to tumors, cells grown using 2-D cell culturing have different proliferation and diffusion rates, cell shapes, and gene expression. To improve in vitro studies, scientists are turning to 3-D cell culturing, which produces microtumors known as spheroids. There are several known techniques for producing spheroids. However, the hanging drop technique is one of the simplest and cheapest methods. Two hanging drop methods, plate and pipette, were evaluated for their ability to produce consistent and transferrable breast cancer (MDA-MB-231) spheroids that are less than 500 µm in diameter. The hanging drop plate method produced the most consistent spheroids. Both the plate and pipette methods produced spheroids larger than 500 µm in diameter. Spheroids were embedded in extracellular matrix at three different collagen concentrations (1.5, 3, 6 mg/mL) at 37 °C. The change in radius over time followed a logarithmic growth pattern. There was no significant difference between spheroids in 1.5 and 6 mg/mL collagen environments. Spheroids were also embedded in extracellular matrix formed at two different temperatures (25 and 37 °C). Spheroids at the lower temperature were significantly larger. A mathematical model was developed to explain the spheroid migration behavior. Key Words: 3-D cell culturing, spheroid, hanging drop, MDA-MB-231
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