, Volume 97, Issue 6, pp 493–497 | Cite as

Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red O

  • J. L. Ramírez-Zacarías
  • F. Castro-Muñozledo
  • W. Kuri-Harcuch


Cultured 3T3-F442A cells differentiate into adipocytes and accumulate lipid droplets in the cytoplasm. When fat cells are stained with Oil red O, the degree of staining seems to be proportional to the extent of cell differentiation. We report here a fast and simple method to quantitate the extent of adipose conversion by staining the accumulated lipid with Oil red O and determining the amount of extracted dye at 510 nm. The results show that Oil red O specifically stains triglycerides and cholesteryl oleate but no other lipids. This technique is a valuable tool for processing large numbers of cell cultures or samples in which adipose differentiation and/or accumulated triglycerides is to be quantitated.


Public Health Lipid Cell Culture Triglyceride Cell Differentiation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ailhaud G (1982) Adipose cell differentiation in culture. Mol Cell Biochem 49:17–31Google Scholar
  2. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein dye binding. Anal Biochem 72:248–253Google Scholar
  3. Fowler SD, Greenspan P (1985) Application of Nile Red, a fluorescent hydrophobic probe for the detection of neutral lipid deposits in tissue sections. J Histochem Cytochem 33:833–836Google Scholar
  4. Green H (1979) Adipose conversion: A program of differentiation. Les Colloques de I'INSERM, Obesity-Cellular and Molecular Aspects 87:15–24Google Scholar
  5. Green H, Kehinde O (1974) Sublines of Mouse 3T3 cells that accumulate lipid. Cell 1:113–116Google Scholar
  6. Green H, Kehinde O (1975) An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion. Cell 5:19–27Google Scholar
  7. Green H, Kehinde O (1976) Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. Cell 7:105–113Google Scholar
  8. Green H, Meuth M (1974) An established pre-adipose cell line and its differentiation in culture. Cell 3:127–133Google Scholar
  9. Humason GL (1972) Animal tissue techniques, 3rd edn. Freeman, San Francisco, pp 307–308Google Scholar
  10. Kasturi R, Joshi VC (1982) Hormonal regulation of stearoyl coenzyme A desaturase activity and lipogenesis during adipose conversion of 3T3-L1 cells. J Biol Chem 257:12224–12230Google Scholar
  11. Kozak LP, Jensen JT (1974) Genetic and developmental control of multiple forms of 1-glycerol-3-phosphate dehydrogenase. J Biol Chem 249:7775–7781Google Scholar
  12. Kruth HS (1984) Filipin-positive, Oil red O-negative particles in atherosclerotic lesions induced by cholesterol feeding. Lab Invest 50:87–93Google Scholar
  13. Kuri-Harcuch W, Green H (1978) Adipose conversion of 3T3 cells depends on a serum factor. Proc Natl Acad Sci USA 75:6107–6109Google Scholar
  14. Kuri-Harcuch W, Wise LS, Green H (1978) Interruption of the adipose conversion of 3T3 cells by biotin deficiency: differentiation without triglyceride accumulation. Cell 14:53–59Google Scholar
  15. Leist DP, Nettleton GS, Feldhoff RC (1986) Determination of lipid loss during aqueous and phase partition fixation using formalin and glutaraldehyde. J Histochem Cytochem 34:437–441Google Scholar
  16. Mays ET, Feldhoff RC, Nettleton GS (1984) Determination of protein loss during aqueous and phase partition fixation using formalin and glutaraldehyde. J Histochem Cytochem 32:1107–1112Google Scholar
  17. Mendoza-Figueroa T, Hernández A, López ML, Kuri-Harcuch W (1988) Intracytoplasmic triglyceride accumulation produced by dexamethasone in adult rat hepatocytes cultivated on 3T3 cells. Toxicology 52:273–286Google Scholar
  18. Morgan TE, Huber GL (1967) Loss of lipid during fixation for electron microscopy. J Cell Biol 32:757–760Google Scholar
  19. Nägele U, Wahlefeld AW, Ziegenhorn J (1985) Triglycerides: UV-methods. In: Bergmeyer J, Graßl M (eds) Methods of enzymatic analysis, vol VIII, 3rd edn. VCH Publishers, Deerfield Beach, Fla., pp 2–12Google Scholar
  20. Nixon T, Green H (1984) Contribution of growth hormone to the adipogenic activity of serum. Endocrinology 114:527–532Google Scholar
  21. Pairault J, Green H (1979) A study of the adipose conversion of suspended 3T3 cells by using glycerophosphate dehydrogenase as differentiation marker. Proc Natl Acad Sci USA 76:5138–5142Google Scholar
  22. Pearse AGE (1968) Histochemistry. Theoretical and applied, vol 1, 3rd edn. Churchill, London, pp 398–446Google Scholar
  23. Spooner PM, Chernick SS, Garrison MM, Scow RO (1979) Development of lipoprotein lipase activity and accumulation of triacylglycerol in differentiating 3T3-L1 adipocytes. Effects of prostaglandin F, 1-methyl-3-isobutylxanthine, prolactin and insulin. J Biol Chem 254:1305–1311Google Scholar
  24. Wahlefeld AW (1974) Triglycerides. Determination after enzymatic hydrolysis. In: Bergmeyer HU (ed) Methods of enzymatic analysis, 2nd English edn. Verlag Chemie International, Deerfield Beach, Fla., pp 1831–1835Google Scholar
  25. Wise LS, Green H (1979) Participation of one isozyme of cytosolic glycerophosphate dehydrogenase in the adipose conversion of 3T3 cells. J Biol Chem 254:273–275Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • J. L. Ramírez-Zacarías
    • 1
  • F. Castro-Muñozledo
    • 1
  • W. Kuri-Harcuch
    • 1
  1. 1.Department of Cell BiologyCentro de Investigación y de Estudios Avanzados del IPNMéxico CityMéxico

Personalised recommendations