Adipocytes differentiated from 3T3L1-MBX fibroblasts were measured with or without an organic extraction method and the Triglyceride-Glo™ Assay. One set of wells was extracted with methanol, transferred to a glass vial and allowed to evaporate to dryness overnight.

No organic extraction was performed on the other set of wells. Both samples were then extracted with the Lysis Solution and triglyceride levels were measured. The triglyceride recovery without organic extraction was higher and less variable over six replicates.

Quantifying triglyceride levels in liver cells is used in liver disorder research for disorders such as non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). Here, human liver microtissues are used to model steatosis of liver cells and quantitating triglyceride levels.

3D InSight™ human Liver Microtissues (MT, InSphero, ~1150 cells/well) were incubated for 3 and 10 days in serum-free medium containing either physiological (LG/LI) or supraphysiological (LG/HI) levels of glucose and insulin and supplementation with either free fatty acids bound to BSA (FFA) or low density lipoprotein plasma fraction (LDL). The microtissues were washed twice in PBS and assayed for total glycerol content according to the standard protocol, as intracellular glycerol content does not contribute significantly to the total glycerol content. Values were plotted as concentration of triglyceride per MT. The data was provided by InSphero, AG, Zurich, Switzerland.