Analysis Of Staining Methods To Identify Neutral Lipid Expression In Human Meibomian Gland Epithelial Cells

Purpose: A critically important step in the differentiation of human meibomian gland epithelial cells (HMGECs) is the intravesicular accumulation of neutral lipids. However, the optimal method to identify such lipids is unclear. The goal of this study was to compare the utility of four different dyes for neutral lipid detection in immortalized (I) HMGECs.Methods: IHMGECs were cultured in a FCS-containing medium for 10 days in the presence or absence of Roxadustat (Roxa), a known-inducer of IHMGEC differentiation (n= 2 wells/treatment/experiment; n= 3 experiments/dye). Cells were then fixed and stained with Oil Red O (ORO), Sudan III (SIII), LipidTox green (LT) or Nile Red (NR) and examined with an Eclipse E800 fluorescent microscope. IHMGECs were evaluated for the number, size and area of stained intracellular lipid vesicles, or the intensity of staining by using bright fields (ORO, SIII) or fluorescence (LT, NR …