Development of in vitro NASH model using hepatocyte spheroid.

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Mouse fibroblast 3T3-L1 was able to be adipogenetic by IBMX and dexamethason under DMEM+FBS condition. We have tried to form spheroid with 3T3-L1. 3T3-L1 did not form spheroid under Williams E medium with insulin and EGF. To solve this problem, we tried two alternative ways. 1. To enhance EGF signaling, we utilized NIH3T3 cells with EGFR. These cells did not form spheroid. 2. To control cell-cell contact, a tumor suppresser gene was introduced into cells. Introduction of RUNX3 cDNA enhanced cell-cell contact of 3T3 cells. E-cadherin expression was increased in RUNX3 transfected cells.increased in RUNX3 transfected cells. While N-cadherin expression was decreased in these cells. RUNX3 introduced 3T3 was able to form spheroid. These cells may be a useful in vitro model for NASH.