title: High content analysis of differentiation and cell death in human adipocytes.
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. © 2013 International Society for Advancement of Cytometry.
authors.affiliation: Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary.
authors.forename: Quang Minh
authors.initials: QM
authors.lastname: Doan-Xuan
authors.forename: Anitta K
authors.initials: AK
authors.lastname: Sarvari
authors.forename: Pamela
authors.initials: P
authors.lastname: Fischer-Posovszky
authors.forename: Martin
authors.initials: M
authors.lastname: Wabitsch
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Balajthy
authors.forename: Laszlo
authors.initials: L
authors.lastname: Fesus
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Bacso
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D050156:Adipogenesis / Q000502:physiology*; D017209:Apoptosis / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D006801:Humans; D007090:Image Interpretation, Computer-Assisted; D047569:Laser Scanning Cytometry / Q000379:methods*
keywords: DNA damage; SGBS; adipocyte; differentiation; high content analysis; laser scanning cytometry; preadipocyte