Differences in the Central Energy Metabolism of Cancer Cells between Conventional 2D and Novel 3D Culture Systems

Abstract

The conventional two-dimensional (2D) culture is available as an in vitro experimental model. However, the culture system reportedly does not recapitulate the in vivo cancer microenvironment. We recently developed a tissueoid cell culture system using Cellbed, which resembles the loose connective tissue in living organisms. The present study performed 2D and three-dimensional (3D) culture using prostate and bladder cancer cell lines and a comprehensive metabolome analysis. Compared to 3D, the 2D culture had significantly lower levels of most metabolites. The 3D culture system did not impair mitochondrial function in the cancer cells and produce energy through the mitochondria simultaneously with aerobic glycolysis. Conversely, ATP production, biomass (nucleotides, amino acids, lipids and NADPH) synthesis and redox balance maintenance were conducted in 3D culture. In contrast, in 2D culture, biomass production was delayed due to the suppression of metabolic activity. The 3D metabolome analysis using the tissueoid cell culture system capable of in vivo cancer cell culture yielded results consistent with previously reported cancer metabolism theories. This system is expected to be an essential experimental tool in a wide range of cancer research fields, especially in preclinical stages while transitioning from in vitro to in vivo.

Assessment of cellular responses in three‐dimensional cell cultures through chemical exchange saturation transfer and 1H MRS

“…The Warburg effect, a cancer hallmark, is the abnormally large percentage of glucose being converted to lactate through glycolysis in cancer cells 48 . This finding agrees with those of studies evaluating metabolites in 2D and 3D cultures through mass spectrometry 59,60 . The large reduction in glucose after DOX treatment may be due to a compensatory mechanism used to maintain adenosine triphosphate (ATP) levels to avoid apoptosis 61 .…”

Cultivation of Cells in a Physiological Plasmax Medium Increases Mitochondrial Respiratory Capacity and Reduces Replication Levels of RNA Viruses

“…In recent years, it has become clear that the investigation of cellular metabolism is affected by cell cultivation conditions. First, the environment which cells find in monolayers in 2D culture is quite different from that in tissue or 3D cultures [ 9 ]. Second, our normoxic condition (21% oxygen) is, rather, superoxic, as in most tissues— except the lung—the oxygen levels are 3.4–6.8% [ 10 , 11 ].…”

Flavonoids Enhance Lipofection Efficiency and Ameliorate Cytotoxicity in Colon26 and HepG2 Cells via Oxidative Stress Regulation

“…Although 2D cell cultures are still used in most studies, 3D cell culture has a broader range of applications as an analysis technology due to features that are closer to the complex in vivo conditions [43], proper cell-cell and cell-extracellular environment interactions [44], high metabolic enzyme expression [45], and possible long-term culture [46]. Therefore, we also investigated the transfection efficiency in 3D cultures of Colon26 cells.…”