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Development of a three‐dimensional in vitro co‐culture model to increase drug selectivity for humans
Author(s) -
Park Sung Bum,
Koh Byumseok,
Jung Won Hoon,
Choi Kyoung Jin,
Na Yoon Ju,
Yoo Hee Min,
Lee Sunray,
Kang Dukjin,
Lee DongMok,
Kim Ki Young
Publication year - 2020
Publication title -
diabetes, obesity and metabolism
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.445
H-Index - 128
eISSN - 1463-1326
pISSN - 1462-8902
DOI - 10.1111/dom.14033
Subject(s) - self healing hydrogels , in vitro , cell culture , in vivo , gelatin , insulin resistance , 3d cell culture , selectivity , chemistry , cell , insulin , western blot , biomedical engineering , biology , biochemistry , medicine , endocrinology , microbiology and biotechnology , genetics , catalysis , organic chemistry , gene
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.