Subsequent to a seven-day period characterized by the accumulation of intracellular lipids, the manifestation of insulin resistance was observed within the spheroids by the 14th day, as indicated by the increased expression levels of phosphoenolpyruvate carboxykinase 1 (PCK1) and pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4), alongside reduced phosphorylation of GSK3β. This evidence concerns the gene PCK1 and Insulin resistance.