Generating Liver Cancer Spheroids for 13C NMR Spectroscopic Studies

The use of spheroids or 3D cell culture method is an important development in pre-clinical studies as spheroids resemble closer morphology and structure to tumors than the conventional 2D cell culture. Herein, 3D cell culture or spheroid results were developed and compared using ultra-low attachment (ULA) plates and the utilization of the hanging drop method. The project focused on two hepatocellular carcinoma liver cancer cell lines HepG2 and HUH7. While the ULA plates were easier to seed, experimental results show there are several advantages to producing spheroids via the hanging drop method. The spheroids collected from the hanging drop method were able to be transferred into regular plasticware designed for 2D culture and placed on an orbital shaker in an incubator for longer cultures. The results of this project will be discussed in light of subsequent 13C NMR spectroscopic studies to probe to aberrant metabolism in cancer cells using the spheroid models.