One-step intracellular triglycerides extraction and quantitative measurement in vitro

J Pharmacol Toxicol Methods. Jul-Aug 2007;56(1):63-6. doi: 10.1016/j.vascn.2007.01.002. Epub 2007 Feb 23.

Abstract

Introduction: Measurement of intracellular triglyceride content is traditionally performed using Oil Red O stain and isopropanol extraction. However, the assay is semi-quantitative and its simplicity needs improving. Therefore, a novel assay is described to address these problems.

Methods: Intracellular triglycerides (TG) were extracted directly by organic solvent Dimethyl Sulfoxide (DMSO) without stain. TG concentration dissolved in DMSO was subsequently measured with a commercial assay kit.

Results: A series of experiments were conducted to verify the validity of the new protocol such as the extraction efficiency of DMSO, linear range of the kit and sensitivity of the assay. Our results suggest that this novel assay may be applied in the measurement of intracellular triglyceride accumulation induced by pre-adipocyte differentiation or compounds such as peroxisome proliferator-activated receptor gamma (PPARgamma) agonists.

Conclusion: The simplicity and efficiency of the method were validated in this study. Our findings show that this novel assay has potential application in the discovery of anti-diabetic compounds or in the research of cell TG metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Dimethyl Sulfoxide
  • Intracellular Space / chemistry
  • Mice
  • Solvents
  • Triglycerides / analysis*
  • Triglycerides / isolation & purification

Substances

  • Solvents
  • Triglycerides
  • Dimethyl Sulfoxide