Quantitation of Adipose Conversion and Triglycerides by Staining Intracytoplasmic Lipids with Oil Red O

Overview

Journal Histochemistry

Specialty Biochemistry

Date 1992 Jul 1

PMID 1385366

Citations 338

Authors

J L Ramirez-Zacarias

F Castro-Munozledo

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Sexually Dimorphic Effects of CYP2B6 in the Development of Fasting-Mediated Steatosis in Mice: Role of the Oxylipin Products 9-HODE and 9-HOTrE.

Eccles-Miller J, Johnson T, Baldwin W Biomedicines. 2025; 13(2).

PMID: 40002708 PMC: 11853041. DOI: 10.3390/biomedicines13020295.

Apolipoprotein B100 acts as a tumor suppressor in ovarian cancer via lipid/ER stress axis-induced blockade of autophagy.

Yin Z, He S, Zhang X, Yu X, Sheng K, Fu T Acta Pharmacol Sin. 2025; .

PMID: 39880927 DOI: 10.1038/s41401-024-01470-x.

Fatty Acid Synthase (FASN) Inhibitors Suppress Metformin-Induced Fat Accumulation and Apoptosis in H4IIE Hepatocellular Carcinoma Cells.

Park D, Boo H Dev Reprod. 2025; 28(4):163-174.

PMID: 39845516 PMC: 11750162. DOI: 10.12717/DR.2024.28.4.163.

Cytotoxicity, Antiadipogenic, Low-Density Lipoprotein Oxidation Inhibitory Activities, and Acute Toxicity Study of Hydroethanolic Leaf and Bark Extracts.

Mba J, Zouheira D, Guetchueng S, Dairou H, Djouonzo P, Ayong L ScientificWorldJournal. 2024; 2024:1732653.

PMID: 39720344 PMC: 11668551. DOI: 10.1155/tswj/1732653.

Effect of Thai Herbal Remedy NL Inhibits Lipid Accumulation on 3T3-L1 Adipocyte Cells.

Warinhomhoun S, Viwatpinyo K, Nokkaew N, Limcharoen T, Ngamdokmai N Adv Pharmacol Pharm Sci. 2024; 2024:2350186.

PMID: 39564001 PMC: 11576086. DOI: 10.1155/2024/2350186.

References

Wise L, Green H . Participation of one isozyme of cytosolic glycerophosphate dehydrogenase in the adipose conversion of 3T3 cells. J Biol Chem. 1979; 254(2):273-5. View

Fowler S, Greenspan P . Application of Nile red, a fluorescent hydrophobic probe, for the detection of neutral lipid deposits in tissue sections: comparison with oil red O. J Histochem Cytochem. 1985; 33(8):833-6. DOI: 10.1177/33.8.4020099. View

Morgan T, Huber G . Loss of lipid during fixation for electron microscopy. J Cell Biol. 1967; 32(3):757-60. PMC: 2107264. DOI: 10.1083/jcb.32.3.757. View

Green H, Kehinde O . An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion. Cell. 1975; 5(1):19-27. DOI: 10.1016/0092-8674(75)90087-2. View

Green H . Adipose conversion of 3T3 cells depends on a serum factor. Proc Natl Acad Sci U S A. 1978; 75(12):6107-9. PMC: 393127. DOI: 10.1073/pnas.75.12.6107. View

Spooner P, CHERNICK S, GARRISON M, SCOW R . Development of lipoprotein lipase activity and accumulation of triacylglycerol in differentiating 3T3-L1 adipocytes. Effects of prostaglandin F2alpha, 1-methyl-3-isobutylxanthine, prolactin, and insulin. J Biol Chem. 1979; 254(4):1305-11. View

Nixon T, Green H . Contribution of growth hormone to the adipogenic activity of serum. Endocrinology. 1984; 114(2):527-32. DOI: 10.1210/endo-114-2-527. View

Wise L, Green H . Interruption of the adipose conversion of 3T3 cells by biotin deficiency: differentiation without triglyceride accumulation. Cell. 1978; 14(1):53-9. DOI: 10.1016/0092-8674(78)90300-8. View

Mendoza-Figueroa T, Hernandez A, De Lourdes Lopez M . Intracytoplasmic triglyceride accumulation produced by dexamethasone in adult rat hepatocytes cultivated on 3T3 cells. Toxicology. 1988; 52(3):273-86. DOI: 10.1016/0300-483x(88)90132-1. View

10.

Pairault J, Green H . A study of the adipose conversion of suspended 3T3 cells by using glycerophosphate dehydrogenase as differentiation marker. Proc Natl Acad Sci U S A. 1979; 76(10):5138-42. PMC: 413095. DOI: 10.1073/pnas.76.10.5138. View

11.

Green H, Kehinde O . Spontaneous heritable changes leading to increased adipose conversion in 3T3 cells. Cell. 1976; 7(1):105-13. DOI: 10.1016/0092-8674(76)90260-9. View

12.

Ailhaud G . Adipose cell differentiation in culture. Mol Cell Biochem. 1982; 49(1):17-31. DOI: 10.1007/BF00230992. View

13.

Green H, Meuth M . An established pre-adipose cell line and its differentiation in culture. Cell. 1974; 3(2):127-33. DOI: 10.1016/0092-8674(74)90116-0. View

14.

Leist D, Nettleton G, Feldhoff R . Determination of lipid loss during aqueous and phase partition fixation using formalin and glutaraldehyde. J Histochem Cytochem. 1986; 34(4):437-41. DOI: 10.1177/34.4.3081623. View

15.

MAYS E, Feldhoff R, Nettleton G . Determination of protein loss during aqueous and phase partition fixation using formalin and glutaraldehyde. J Histochem Cytochem. 1984; 32(10):1107-12. DOI: 10.1177/32.10.6434629. View

16.

Kruth H . Filipin-positive, oil red O-negative particles in atherosclerotic lesions induced by cholesterol feeding. Lab Invest. 1984; 50(1):87-93. View

17.

Kozak L, Jensen J . Genetic and developmental control of multiple forms of L-glycerol 3-phosphate dehydrogenase. J Biol Chem. 1974; 249(24):7775-81. View

18.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

19.

Kasturi R, Joshi V . Hormonal regulation of stearoyl coenzyme A desaturase activity and lipogenesis during adipose conversion of 3T3-L1 cells. J Biol Chem. 1982; 257(20):12224-30. View