Progression of metabolic dysfunction-associated steatotic liver disease leads to insulin resistance, a condition driven by the induction of specific hepatokines. We previously developed an in vitro steatotic liver model, PXB-cells LA (Lipid Analysis), derived from fresh human hepatocytes (PXB-cells^®) isolated from humanized mouse livers. In the present study, we investigated its utility as a screening system for insulin-sensitizing agents. Our results demonstrated that metformin and rosiglitazone, well-known treatments for type 2 diabetes, reduced the expression of the prodiabetic hepatokines leukocyte cell-derived chemotaxin 2 and fetuin-A in PXB-cells LA. Furthermore, while fibroblast growth factor 21 enhances systemic insulin sensitivity, both agents upregulated fibroblast growth factor 21 production at both the mRNA and protein levels in this model.