Prolonged treatment with 3-isobutyl-1-methylxanthine improves the efficiency of differentiating 3T3-L1 cells into adipocytes

Yongjie Hua; Shanshan Ke; Yao Wang; David M Irwin; Shuyi Zhang; Zhe Wang

doi:10.1016/j.ab.2016.05.007

Prolonged treatment with 3-isobutyl-1-methylxanthine improves the efficiency of differentiating 3T3-L1 cells into adipocytes

Anal Biochem. 2016 Aug 15:507:18-20. doi: 10.1016/j.ab.2016.05.007. Epub 2016 May 19.

Authors

Yongjie Hua¹, Shanshan Ke², Yao Wang², David M Irwin³, Shuyi Zhang⁴, Zhe Wang⁵

Affiliations

¹ State Key Laboratory of Estuarine and Coastal Research, Institute of Estuarine and Coastal Research, East China Normal University, Shanghai 200062, China.
² Institute of Molecular Ecology and Evolution, East China Normal University, Shanghai, 200062, China.
³ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, M5S 1A1, Canada.
⁴ State Key Laboratory of Estuarine and Coastal Research, Institute of Estuarine and Coastal Research, East China Normal University, Shanghai 200062, China; Key Laboratory of Zoonosis of Liaoning Province, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110866, China.
⁵ State Key Laboratory of Estuarine and Coastal Research, Institute of Estuarine and Coastal Research, East China Normal University, Shanghai 200062, China; Key Laboratory of Zoonosis of Liaoning Province, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110866, China. Electronic address: [email protected].

PMID: 27210514
DOI: 10.1016/j.ab.2016.05.007

Abstract

Until now, the low efficiency of current protocols or kits for the differentiation of 3T3-L1 preadipocytes makes it difficult to continue the studies of the cellular and molecular mechanisms in adipocytes. Here we present a productive and highly efficient protocol for the differentiation of 3T3-L1 cells that uses a prolonged treatment with 3-isobutyl-1-methylxanthine (IBMX) during the differentiated process. 3T3-L1 cells of unknown passage +3 and unknown passage +7 treated with a prolonged exposure to IBMX showed significantly increased differentiation efficiency by day 15, in contrast to low levels of differentiation seen with protocols that lacked additional IBMX.

Keywords: 3T3-L1 cell; Adipocyte differentiation; C/EBPβ; IBMX.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

1-Methyl-3-isobutylxanthine / administration & dosage
1-Methyl-3-isobutylxanthine / pharmacology*
3T3-L1 Cells
Adipocytes / cytology*
Adipocytes / drug effects*
Animals
Cell Differentiation / drug effects*
Cell Proliferation / drug effects
Dose-Response Relationship, Drug
Mice
Structure-Activity Relationship
Time Factors

Substances

1-Methyl-3-isobutylxanthine