Quantification of sterol-specific response in human macrophages using automated imaged-based analysis

Deborah L. Gater, Namareq Widatalla, Kinza Islam, Maryam Alraeesi, Jeremy Teo, Yanthe E. Pearson

Research output: Contribution to journalArticle

Abstract

Background: The transformation of normal macrophage cells into lipid-laden foam cells is an important step in the progression of atherosclerosis. One major contributor to foam cell formation in vivo is the intracellular accumulation of cholesterol. Methods: Here, we report the effects of various combinations of low-density lipoprotein, sterols, lipids and other factors on human macrophages, using an automated image analysis program to quantitatively compare single cell properties, such as cell size and lipid content, in different conditions. Results: We observed that the addition of cholesterol caused an increase in average cell lipid content across a range of conditions. All of the sterol-lipid mixtures examined were capable of inducing increases in average cell lipid content, with variations in the distribution of the response, in cytotoxicity and in how the sterol-lipid combination interacted with other activating factors. For example, cholesterol and lipopolysaccharide acted synergistically to increase cell lipid content while also increasing cell survival compared with the addition of lipopolysaccharide alone. Additionally, ergosterol and cholesteryl hemisuccinate caused similar increases in lipid content but also exhibited considerably greater cytotoxicity than cholesterol. Conclusions: The use of automated image analysis enables us to assess not only changes in average cell size and content, but also to rapidly and automatically compare population distributions based on simple fluorescence images. Our observations add to increasing understanding of the complex and multifactorial nature of foam-cell formation and provide a novel approach to assessing the heterogeneity of macrophage response to a variety of factors.

Original languageEnglish (US)
Article number242
JournalLipids in Health and Disease
Volume16
Issue number1
DOIs
StatePublished - Dec 13 2017

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Macrophages
Sterols
Lipids
Foam Cells
Cholesterol
Foams
Cytotoxicity
Cell Size
Image analysis
Lipopolysaccharides
Population distribution
Ergosterol
LDL Lipoproteins
Cell Survival
Atherosclerosis
Fluorescence
Cells
Demography

Keywords

  • Cholesterol
  • Ergosterol
  • Foam cell
  • Image processing
  • Lipid droplet
  • Lysophosphatidylcholine
  • THP-1
  • Vesicle
  • Watershedding

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Quantification of sterol-specific response in human macrophages using automated imaged-based analysis. / Gater, Deborah L.; Widatalla, Namareq; Islam, Kinza; Alraeesi, Maryam; Teo, Jeremy; Pearson, Yanthe E.

In: Lipids in Health and Disease, Vol. 16, No. 1, 242, 13.12.2017.

Research output: Contribution to journalArticle

Gater, Deborah L. ; Widatalla, Namareq ; Islam, Kinza ; Alraeesi, Maryam ; Teo, Jeremy ; Pearson, Yanthe E. / Quantification of sterol-specific response in human macrophages using automated imaged-based analysis. In: Lipids in Health and Disease. 2017 ; Vol. 16, No. 1.
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