Development of in vitro three‐dimensional co‐culture system for metabolic syndrome therapeutic agents

Abstract

Aims There are many obstacles to overcome in the development of new drugs for metabolic diseases, including efficacy and toxicity problems in later stages of drug development. To overcome these problems and predict efficacy and toxicity in early stages, we constructed a new model of insulin resistance in terms of communication between 3T3‐L1 adipocytes and RAW264.7 macrophages by three‐dimensional (3D) culture. Results In this study, results focused on the functional resemblance between 3D co–culture of adipocytes and macrophages and adipose tissue in diabetic mice. The 3D mono–culture preadipocytes showed good cell viability and induced cell differentiation to adipocytes, without cell confluence or cell–cell contact and interaction. The 3D co‐cultured preadipocytes with RAW264.7 macrophages induced greater insulin resistance than two‐dimensional and 3D mono‐cultured adipocytes. Additionally, we demonstrated that 3D co‐culture model had functional metabolic similarity to adipose tissue in diabetic mice. We utilized this 3D co‐culture system to screen PPARγ antagonists that might have potential as therapeutic agents for diabetes as demonstrated by an in vivo assay. Conclusion This in vitro 3D co‐culture system could serve as a next‐generation platform to accelerate the development of therapeutics for metabolic diseases.

iTRAQ-Based Quantitative Proteomic Comparison of 2D and 3D Adipocyte Cell Models Co-cultured with Macrophages Using Online 2D-nanoLC-ESI-MS/MS

“…Thus, attempts to develop in vitro 3D cell culture systems have been undertaken. In our previous study 30 , we demonstrated that the co-culture of adipocytes and macrophages in a 3D cell culture system results in changes in lipid and glucose metabolism, which is similar to that of the effect of GW9662 on insulin resistance in adipose tissue in diabetic mice. However, there is no clear reason why the insulin sensitivity in adipocytes induced by 3D co-culturing models with macrophages was shown to be consistent with the response observed in vivo in adipose tissue.…”

“…Both physical and functional aspects of adipocytes that are co-cultured with the other cell types during differentiation and enlargement differed significantly in terms of cell morphology and cytokine expression compared to that of mono-cultured adipocytes 28 . In our previous study 30 , we also found that the expression levels of the metabolic pathway-related proteins from 3D co-cultured adipocytes with macrophages changed distinctively compared to 2D and 3D mono-cultured adipocytes. To clarify the metabolic differences between 2D and 3D mono/co-culture models, however, a comprehensive proteomic analysis of adipocyte proteome is required.…”

“…In our previous study, we developed a 3D cell printing system to assemble 3D-dispensing structures 30 . Briefly, this system consists of an x-y-z stage, a dispenser, a syringe nozzle, a compression controller and a computer system.…”

On the Road to Accurate Biomarkers for Cardiometabolic Diseases by Integrating Precision and Gender Medicine Approaches

“…This could also facilitate their complex management and therapy, the most important objectives of clinical community. Certainly, the use of more appropriate study models, such as the innovative three-dimensional (3D) cardiometabolic tissue constructs, can help in the integration of multi-omics approaches [37,38] (see Figure 2). The 3D systems attempt to address limitations of the conventional two-dimensional (2D) in vitro models, such as monoculture cellular assays, as well as in vivo animal models [37,38,39].…”

“…Certainly, the use of more appropriate study models, such as the innovative three-dimensional (3D) cardiometabolic tissue constructs, can help in the integration of multi-omics approaches [37,38] (see Figure 2). The 3D systems attempt to address limitations of the conventional two-dimensional (2D) in vitro models, such as monoculture cellular assays, as well as in vivo animal models [37,38,39]. The 2D in vitro models have limitations in the simulation of CMD pathophysiology because of the high degree of complexity in the structure and function of the cardiometabolic tissues [37,38].…”