Speaker
Description
Introduction:
Petridish-based 2-dimensional (2D) cell culture models are currently used as the first step to test the efficacy of anticancer drugs, but they don’t accurately mimic tumor biology and microenvironment. Moving to 3D models, one of the major concerns is how to assess the drug efficacy. Cell viability is an essential parameter to be monitored to measure cell growth and to assess the effectiveness of a drug candidate in cancer research. Several cell viability assays are commonly used but these assays are typically used in low cell density 2D cultures and may not produce the same results in 3D cultures with high cell density and added matrix components. Our aim is to develop a realistic preclinical 3D brain tumour model from native tissue and evaluate the accuracy of our 3D model to reflect a realistic drug response.
Methodology:
3D brain tumour model were developed by decellularizing animal brain tissue followed by seeding with U251 cells. Identical U251 cell numbers were plated in 2D and were mixed with Matrigel (a common 3D cell culture platform) to form a 3D culture model. We compared the results of three different viability assays (MTT, Alamar Blue, ATP) on three different culture models and calibrated the assay output versus known cell numbers by counting and DNA quantification.
Results:
In all of our selected cell viability assays, the assay output was higher in 2D samples compared to matrigel samples containing the same number of cells. In our novel 3D brain tumour model, ATP assay and Alamar Blue assay showed the most similarity to the DNA assay. MTT showed the most difference in assay read outs in 2D vs Matrigel, the same result reflected in the scaffolds.
Conclusion:
The results suggested the 3D brain tumour model performed better than Matrigel based-model in terms of accuracy of assay readouts (ATP and Alamar Blue). MTT was observed to not measure the viability of the cancer cells accurately for all 3D models. Not all protocols for viability assays that are validated for 2D cell culture are directly applicable to 3D models. Further optimization and calibration are required for 3D cell culture.
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