Prolonged Induction Activates Cebpα Independent Adipogenesis in NIH/3T3 Cells
Journal
PLoS ONE
Journal Volume
8
Journal Issue
1
Pages
e51459
Date Issued
2013
Author(s)
Abstract
Background: 3T3-L1 cells are widely used to study adipogenesis and insulin response. Their adipogenic potential decreases with time in the culture. Expressing exogenous genes in 3T3-L1 cells can be challenging. This work tries to establish and characterize an alternative model of cultured adipocytes that is easier to work with than the 3T3-L1 cells. Methodology/Principal Findings: Induced cells were identified as adipocytes based on the following three characteristics: (1) Accumulation of triglyceride droplets as demonstrated by oil red O stain. (2) Transport rate of 2-deoxyglucose increased after insulin stimulation. (3) Expression of fat specific genes such as Fabp4 (aP2), Slc2a4 (Glut4) and Pparg (PPARγ). Among the cell lines induced under different conditions in this study, only NIH/3T3 cells differentiated into adipocytes after prolonged incubation in 3T3-L1 induction medium containing 20% instead of 10% fetal bovine serum. Rosiglitazone added to the induction medium shortened the incubation period from 14 to 7 days. The PI3K/AKT pathway showed similar changes upon insulin stimulation in these two adipocytes. C/EBPα mRNA was barely detectable in NIH/3T3 adipocytes. NIH/3T3 adipocytes induced in the presence of rosiglitazone showed higher 2-deoxyglucose transport rate after insulin stimulation, expressed less Agt (angiotensinogen) and more PPARγ. Knockdown of C/EBPα using shRNA blocked 3T3-L1 but not NIH/3T3 cell differentiation. Mouse adipose tissues from various anatomical locations showed comparable levels of C/EBPα mRNA. Conclusions/Significance: NIH/3T3 cells were capable of differentiating into adipocytes without genetic engineering. They were an adipocyte model that did not require the reciprocal activation between C/EBPα and PPARγ to differentiate. Future studies in the C/EBPα independent pathways leading to insulin responsiveness may reveal new targets to diabetes treatment. ? 2013 Shao et al.
SDGs
Other Subjects
angiotensinogen; CCAAT enhancer binding protein alpha; deoxyglucose; fat droplet; fatty acid binding protein 4; glucose transporter 4; insulin; messenger RNA; peroxisome proliferator activated receptor gamma; phosphatidylinositol 3 kinase; protein kinase B; rosiglitazone; short hairpin RNA; triacylglycerol; adipocyte; adipogenesis; adipose tissue; animal cell; animal tissue; article; carbohydrate transport; cell differentiation; cell strain 3T3; controlled study; culture medium; diabetes mellitus; Fabp4 gene; gene expression; gene silencing; genetic engineering; incubation time; lipid storage; mouse; nonhuman; PPARG gene; protein induction; signal transduction; SLC2A4 gene; 3T3-L1 Cells; Adipocytes; Adipogenesis; Animals; Biological Transport; Blotting, Western; Cattle; CCAAT-Enhancer-Binding Protein-alpha; Cell Differentiation; Culture Media; Deoxyglucose; Fatty Acid-Binding Proteins; Gene Expression; Hypoglycemic Agents; Insulin; Mice; Mice, Inbred C57BL; NIH 3T3 Cells; PPAR gamma; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Serum; Thiazolidinediones; Time Factors; Triglycerides; Bovinae
Type
journal article