Cytotechnology

, Volume 46, Issue 1, pp 49–56 | Cite as

Oil red-O stains non-adipogenic cells: a precautionary note

  • Adrienne D. Kinkel
  • Melinda  E. Fernyhough
  • Deri L. Helterline
  • Janet L. Vierck
  • Karen S. Oberg
  • Tyler J. Vance
  • Gary J. Hausman
  • Rodney A. Hill
  • Michael V. Dodson
Article

Abstract

Bovine adipofibroblasts, 3T3-L1 cells, L-6 myogenic cells, and sheep satellite cells were allowed to proliferate for 48 h. Oil red-O (ORO) was dissolved in three different solvents isopropanol, propylene glycol and triethyl phosphate. At 48 h, the proliferative cultures were stained with the three stains. ORO stain prepared in both propylene glycol and triethyl phosphate resulted in bright red droplets appearing in all cultures, whereas ORO dissolved in isopropanol was not taken up by any of the cells. These data suggest that certain preparations of ORO may stain cells in non-adipogenic lineages as well as undifferentiated pre-adipocytes. Caution must be exercised when choosing solvents for ORO in differentiation studies using cells of the fat/adipose lineage.

Keywords

Adipocyte Adipofibroblasts Cell culture Oil red-O Satellite cell Staining 

Abbreviation

DMEM

Dulbecco’s modified Eagle’s medium

FBS

Fetal bovine serum

ORO

Oil red-O

PBS

Phosphate-buffered saline, pH 7.08

PEG

Propylene glycol

PSG

Pig skin gelatin

SC

Satellite cell

TEP

Triethyl phosphate

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adebonojo, F.O. 1975Studies on human adipose cells in culture: relation of cell size and cell multiplication to donor ageYale J. Biol. Med.48916PubMedGoogle Scholar
  2. Dodson, M.V., McFarland, D.C., Martin, E.L., Brannon, M.A. 1986Isolation of satellite cells from ovine skeletal musclesJ. Tissue Cult. Meth.10233237CrossRefGoogle Scholar
  3. Feldman, A., Dapson, R. 1974Relative effectiveness of various solvents for oil red OMed. Lab. Tech.31335341Google Scholar
  4. Fukumoto, S., Fumimoto, T. 2002Deformation of lipid droplets in fixed samplesHistochem. Cell Biol.118423428CrossRefPubMedGoogle Scholar
  5. Gamou, S., Shimizu, Y., Shimizu, N. 1990

    Adipocytes

    Pollard, J.W.Walker, J.M. eds. Methods in Molecular Biology, Vol. 5Humana PressNew Jersey197207
    Google Scholar
  6. Johnson, P.R., Francendese, A.A. 1985Cellular regulation of adipose tissue growthJ. Anim. Sci.615775Google Scholar
  7. Koopman, R., Schaart, G., Hesselink, M.K.C. 2001Optimisation of oil red O staining permits combination with immunofluorescence and automated quantification of lipidsHistochem. Cell Biol.1166368PubMedGoogle Scholar
  8. Lillie, R.D., Ashburn, L.L. 1943Supersaturated solutions of fat stains in dilute isopropanol for demonstration of acute fatty degeneration not shown by Herxheimer’s techniqueArch. Pathol.36432440Google Scholar
  9. Putt, F.A. 1972Manual of Histopathological Staining MethodsJohn WileyNew York194196Google Scholar
  10. Ramìrez-Zacarìas, J.L., Castro-Munozledo, F., Kuri-Harchuch, W. 1992Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red OHistochem. J.97493497CrossRefGoogle Scholar
  11. Vierck, J.L., Dal Porto, D., Dodson, M.V. 2001Induction of preadipocyte differentiation by a defined treatment medium without DMIBasic Appl. Myol.1199104Google Scholar
  12. Vierck, J.L., McNamara, J.P., Dodson, M.V. 1996Two alternative procedures to isolate adipofibroblasts from sheep skeletal muscleMeth. Cell Sci.18309314CrossRefGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Adrienne D. Kinkel
    • 1
  • Melinda  E. Fernyhough
    • 1
  • Deri L. Helterline
    • 1
  • Janet L. Vierck
    • 1
  • Karen S. Oberg
    • 1
  • Tyler J. Vance
    • 1
  • Gary J. Hausman
    • 2
  • Rodney A. Hill
    • 3
  • Michael V. Dodson
    • 1
  1. 1.Muscle Biology Laboratory, Department of Animal Sciences Washington State UniversityPullmanUSA
  2. 2.Richard B. Russell Agricultural Research CenterUSDA-ARSAthensUSA
  3. 3.Department of Animal and Veterinary ScienceUniversity of IdahoMoscowUSA

Personalised recommendations