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

Ryo Ikari,Shigehisa Kubota,Takahisa Nakayama,Masayuki Nagasawa,Ken-Ichi Mukaisho,Akihiro Kawauchi,Shoko Murakami,Ryoji P Kushima,Susumu Kageyama,Naoko Taniura,Hiroyuki Tanaka

doi:10.3390/ijms22041805

Ryo Ikari, Shigehisa Kubota + Show 9 more

Open Access

https://doi.org/10.3390/ijms22041805

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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.

Highlights

Cancer research, such as the development of anticancer drugs, requires the development of feasible experimental models of organisms that are easy-to-use and cost-effective.At present, a conventional easy-to-use in vitro experimental model of a two-dimensional (2D) culture method is available
11% of in vivo drug resistance studies using the 2D culture system reach the final phase of demonstrating sufficient efficacy and safety [7], whereas those using the three-dimensional (3D) culture system appear to better simulate the organism’s environment [8]. 3D culture systems using spheroids, extracellular matrices, systems for generating cancer organoids and cancer-tissue-originated spheroid methods have recently been reported, showing the fast development of 3D culture [9]
Highly differentiated LNCaP cells showed nodular growth, whereas PC-3 cells with poor differentiation showed a mixture of cancer cells that formed small agglomerates and those that exist alone

Summary

Introduction

Cancer research, such as the development of anticancer drugs, requires the development of feasible experimental models of organisms that are easy-to-use and cost-effective. A conventional easy-to-use in vitro experimental model of a two-dimensional (2D) culture method is available. Most studies utilizing this experimental method have failed to demonstrate sufficient safety and efficacy of test drugs in the preclinical stage [1,2,3,4,5,6]. 11% (only 5% in the field of oncology) of in vivo drug resistance studies using the 2D culture system reach the final phase of demonstrating sufficient efficacy and safety [7], whereas those using the three-dimensional (3D) culture system appear to better simulate the organism’s environment [8]. Previous 3D culture studies of various cancer cell lines, such as colorectal cancer, Barrett0 s adenocarcinoma, gastroesophageal junction cancer, gastric cancer and tongue cancer, have shown that 3D culture systems allow good morphological observations resembling in vivo conditions [10]

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