Development of a three‐dimensional in vitro co‐culture model to increase drug selectivity for humans. Issue 8 (16th April 2020)

• Record Type: Journal Article
• Authors: Park, Sung Bum; Koh, Byumseok; Jung, Won Hoon; Choi, Kyoung Jin; Na, Yoon Ju; Yoo, Hee Min; Lee, Sunray; Kang, Dukjin; Lee, Dong‐Mok; Kim, Ki Young
• Is Part Of: Diabetes, obesity & metabolism. Volume 22:Issue 8(2020)
• Page Start: 1302
• Page End: 1315
• Publication Date: 2020-04-16
• Subjects: antidiabetic drug -- antiobesity drug -- drug development -- type 2 diabetes; Diabetes -- Periodicals; Obesity -- Periodicals; Metabolism -- Disorders -- Periodicals; Clinical pharmacology -- Periodicals; 616.462
• DOI: 10.1111/dom.14033
• Languages: English
• ISSNs: 1462-8902

Abstract

Abstract: Aim: Insulin resistance is a metabolic state where insulin sensitivity is lower than normal condition and strongly related to type 2 diabetes. However, an in vitro model mimicking insulin resistance is rare and thus screening drugs for insulin resistance severely depends on an in vivo model. Here, to increase anti‐diabetic drug selectivity for humans, 3D ADMSCs and macrophages were co‐cultured with in‐house fabricated co‐culture plates. Material and methods: 3D co‐culture plates were designed to load ADMSCs and RAW264.7 cells containing hydrogels in separate wells while allowing cell‐cell interaction with co‐culturing media. Hydrogels were constructed using a 3D cell‐printing system containing 20 mg/ml alginate, 0.5 mg/ml gelatin and 0.5 mg/ml type I collagen. Cells containing hydrogels in 3D co‐culture plates were incubated for 10 min to allow stabilization before the experiment. 3D co‐culture plates were incubated with the CaCl2 solution for 5 min to complete the cross linking of alginate hydrogel. Cells in 3D co‐culture plates were cultured for up to 12 days depending on the experiment and wells containing adipocytes and macrophages were separated and used for assays. Results: KR‐1, KR‐2 and KR‐3 compounds were applied during differentiation (12 days) in 3D co‐cultured mouse 3T3‐L1 adipocytes and 3D co‐cultured human ADMSCs. Glucose uptake assay using 2‐DG6P and 2‐NBDG and western blot analysis were performed to investigate changes of insulin resistance in the 3D co‐cultured model for interspecies selectivity of drug screening. KR‐1 (mouse potent enantiomer) and KR‐3 (racemic mixture) showed improvement of 2‐DG and 2‐NBDG uptake compared with KR‐2 (human potent enantiomer) in 3D co‐cultured 3T3‐L1 adipocytes. In connection with insulin resistance in a 3D 3T3‐L1 co‐cultured model, KR‐1 and KR‐3 showed improvement of insulin sensitivity compared to KR‐2 by markedly increasing GLUT4 expression. In contrast to the result of 3D co‐cultured 3T3‐L1 adipocytes, KR‐1 failed to significantly improve 2‐DG and 2‐NBDG uptake in 3D co‐cultured ADMSC adipocytes. Results of 2‐NBDG accumulation and western blot analysis also showed that KR‐2 and KR‐3 improved insulin sensitivity relatively better than KR‐1. Conclusions: Our 3D co‐culture model with/without 3D co‐culture plates can successfully mimic insulin resistance while allowing investigation of the effects of anti‐obesity or anti‐diabetic drugs on human or mouse co‐culturing cell type. This 3D co‐culture system may accelerate screening of drugs for insulin resistance depending on species.