High content analysis of differentiation and cell death in human adipocytes

Quang Minh Doan-Xuan, Anitta K. Sarvari, Pamela Fischer-Posovszky, Martin Wabitsch, Zoltan Balajthy, Laszlo Fesus, Zsolt Bacso

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Understanding adipocyte biology and its homeostasis is in the focus of current obesity research. We aimed to introduce a high-content analysis procedure for directly visualizing and quantifying adipogenesis and adipoapoptosis by laser scanning cytometry (LSC) in a large population of cell. Slide-based image cytometry and image processing algorithms were used and optimized for high-throughput analysis of differentiating cells and apoptotic processes in cell culture at high confluence. Both preadipocytes and adipocytes were simultaneously scrutinized for lipid accumulation, texture properties, nuclear condensation, and DNA fragmentation. Adipocyte commitment was found after incubation in adipogenic medium for 3 days identified by lipid droplet formation and increased light absorption, while terminal differentiation of adipocytes occurred throughout day 9-14 with characteristic nuclear shrinkage, eccentric nuclei localization, chromatin condensation, and massive lipid deposition. Preadipocytes were shown to be more prone to tumor necrosis factor alpha (TNFα)-induced apoptosis compared to mature adipocytes. Importantly, spontaneous DNA fragmentation was observed at early stage when adipocyte commitment occurs. This DNA damage was independent from either spontaneous or induced apoptosis and probably was part of the differentiation program.

Original languageEnglish
Pages (from-to)933-943
Number of pages11
JournalCytometry Part A
Volume83
Issue number10
DOIs
Publication statusPublished - Oct 1 2013

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Keywords

  • Adipocyte
  • DNA damage
  • Differentiation
  • High content analysis
  • Laser scanning cytometry
  • Preadipocyte
  • SGBS

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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