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Evaluation of adipocyte apoptosis by laser scanning cytometry

International Journal of Obesity volume 28, pages 1535–1540 (2004)Cite this article

Abstract

OBJECTIVE: Adipocyte apoptosis plays an important role in adipose tissue homeostasis and can be altered under a variety of physiological and pathological conditions. This study was carried out to determine whether laser scanning cytometry (LSC) can be used to measure changes in apoptosis of adipocytes over time.

DESIGN: LSC was used to investigate adipocyte apoptosis induced by tumor necrosis factor-alpha (TNF-α), a cytokine that is associated with obesity and insulin resistance. LSC, a slide-based solid phase cytofluorometer, provides quantitative flow fluorescence data together with morphological information for apoptotic detection. Both 3T3-L1 cells and rat adipocytes from primary cell culture were incubated with 0 or 25 nM TNF-α for up to 24 h. Both the FITC-conjugated annexin V/propidium iodide assay and the TUNEL assay were used to distinguish cells with apoptotic characteristics from nonapoptotic cells.

RESULTS: Apoptosis did not increase over time in the absence of TNF-α for both 3T3-L1 cells and rat primary adipocytes. For both 3T3-L1 cells and rat primary adipocytes, a significant increase in the percentage of apoptotic cells was observed by 3–4 h incubation with TNF-α (P<0.05). By 24 h, more than 50% of cells incubated with TNF-α were apoptotic (P<0.001). This process was also associated with morphological changes typical of adipocytes undergoing apoptosis. By estimating the percentage of cell subpopulations after different times of incubation with TNF-α, we were able to develop grading parameters, based on the adipose apoptotic measurements.

CONCLUSION: With morphological information, LSC can be a useful tool to evaluate adipocyte apoptosis.

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Acknowledgements

This work was supported in part by the Georgia Research Alliance Eminent Scholar endowment held by CAB.

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Authors and Affiliations

  1. Department of Animal and Dairy Science, University of Georgia, Athens, GA, USA

    J Lin, K A Page, M A Della-Fera & C A Baile

  2. Department of Foods and Nutrition, University of Georgia, Athens, GA, USA

    C A Baile

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  1. J Lin
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  2. K A Page
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Correspondence to C A Baile.

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Lin, J., Page, K., Della-Fera, M. et al. Evaluation of adipocyte apoptosis by laser scanning cytometry. Int J Obes 28, 1535–1540 (2004). https://doi.org/10.1038/sj.ijo.0802777

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