Image-Based High-Throughput Quantification of Cellular Fat Accumulation

A number of biochemical methods are available for measuring fat accumulation in cell culture. The authors report a simple image-based method for measuring fat accumulation in adipocytes using a combination of high-throughput brightfield microscopy and image analysis, which was validated biochemically using Oil-Red-O. The quickest and most accurate method of analysis was one based on thresholding brightfield images and determining the area of fat droplets per image. Thus, the authors have developed a simple high-throughput, label-free method for measuring fat accumulation that is applicable to any cell or tissue type where fat droplets are visible under light microscopy. (Journal of Biomolecular Screening 2007:999-1005)

[1]  B. Spiegelman,et al.  Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. , 1994, Cell.

[2]  Christer S. Ejsing,et al.  Polyene-lipids: A new tool to image lipids , 2005, Nature Methods.

[3]  M. Lane,et al.  CCAAT/enhancer-binding protein β is required for mitotic clonal expansion during adipogenesis , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Thomas D. Y. Chung,et al.  A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays , 1999, Journal of biomolecular screening.

[5]  Shizuka Hirai,et al.  Activin A inhibits differentiation of 3T3-L1 preadipocyte , 2005, Molecular and Cellular Endocrinology.

[6]  Hongyun Yan,et al.  Role of cyclooxygenases COX-1 and COX-2 in modulating adipogenesis in 3T3-L1 cells Published, JLR Papers in Press, November 4, 2002. DOI 10.1194/jlr.M200357-JLR200 , 2003, Journal of Lipid Research.

[7]  B. Spiegelman,et al.  Transdifferentiation of myoblasts by the adipogenic transcription factors PPAR gamma and C/EBP alpha. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Lehmann,et al.  A prostaglandin J2 metabolite binds peroxisome proliferator-activated receptor γ and promotes adipocyte differentiation , 1995, Cell.

[9]  J. L. Ramírez-Zacarías,et al.  Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red O , 1992, Histochemistry.

[10]  H. Green,et al.  An established preadipose cell line and its differentiation in culture II. Factors affecting the adipose conversion , 1975, Cell.

[11]  M. Lane,et al.  Mitotic clonal expansion: A synchronous process required for adipogenesis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[12]  H. Green,et al.  Participation of one isozyme of cytosolic glycerophosphate dehydrogenase in the adipose conversion of 3T3 cells. , 1979, The Journal of biological chemistry.

[13]  L. Fajas,et al.  Cyclin D3 Promotes Adipogenesis through Activation of Peroxisome Proliferator-Activated Receptor γ , 2005, Molecular and Cellular Biology.

[14]  H. Green,et al.  Sublines of mouse 3T3 cells that accumulate lipid , 1974 .

[15]  B. Spiegelman,et al.  Molecular regulation of adipogenesis. , 2000, Annual review of cell and developmental biology.

[16]  R. Morrison,et al.  Role of PPARγ in Regulating a Cascade Expression of Cyclin-dependent Kinase Inhibitors, p18(INK4c) and p21(Waf1/Cip1), during Adipogenesis* , 1999, The Journal of Biological Chemistry.

[17]  B. Spiegelman,et al.  Stimulation of adipogenesis in fibroblasts by PPARγ2, a lipid-activated transcription factor , 1994, Cell.