Glioblastoma Cancer Neurosphere Formation In-Vitro using a Non-Serum Stem-Cell Media

Mackenzie Miller*, Sonikpreet Aulakh, and Adrienne Williams

Leonard C. Nelson College of Engineering and Sciences, Department of Biology, West Virginia University Institute of Technology, Beckley, WV 25801

Presentation Category: Oral-Science & Technology (Oral Presentation #36)

Student’s Major: Biology

Glioblastoma multiforme (GBM) is one of the most lethal cancers in the world. Almost 12,500 people each year are diagnosed with GBM with only about 35%-survival past one year. This raises a need for research into glioblastoma. However, traditional laboratory techniques used to maintain glioblastomas fail to replicate their real-life 3-dimensional (3D) nature, which makes it difficult to find effective treatment options. The purpose of this study was to find an effective laboratory growth condition for glioma cancer cells to form neurospheres (small neural cell clusters) that maintain original tumor characteristics. To do this, glioma cancer cells (LN229) were divided, resuspended in an appropriate growth medium, and transferred to 60mm plastic dishes (replicates of 4), and monitored every two days for up to 10 days maintained at 37°C with 5% carbon dioxide. Cell growth and neurosphere formation were monitored using a colorimetric cell viability assay and light microscopy. For each experiment (n = 2), the control condition contained typical blood serum-based growth medium compared to our experimental non-serum-based growth medium that contained essential stem-cell growth factors. Our hypothesis was that non-serum stem-cell media would form neurospheres most successfully. Non-serum-based media demonstrated significantly increased numbers of neurosphere formation (p < 0.05) and neurosphere diameter (p < 0.05) compared to the control condition. We found no significant difference in cell viability over the course of the experiments. Therefore, using the non-serum stem-cell media enhanced effectiveness of studying 3D glioblastoma growth in the laboratory setting and will be used for future treatment experimentation.

Funding:

Program/mechanism supporting research/creative efforts: a WVU 497-level course