LDL cholesterol uptake assay using live cell imaging analysis with cell health monitoring

Portia Ritter, Keyvan Yousefi, Juliana Ramirez, Derek M Dykxhoorn, Armando J Mendez, Lina Shehadeh

Research output: Contribution to journalArticle

Abstract

The regulation of LDL cholesterol uptake through LDLR-mediated endocytosis is an important area of study in various major pathologies including metabolic disorder, cardiovascular disease, and kidney disease. Currently, there is no available method to assess LDL uptake while simultaneously monitoring for health of the cells. The current study presents a protocol, using a live cell imaging analysis system, to acquire serial measurements of LDL influx with concurrent monitoring for cell health. This novel technique is tested in three human cell lines (hepatic, renal tubular epithelial, and coronary artery endothelial cells) over a four-hour time course. Moreover, the sensitivity of this technique is validated with well-known LDL uptake inhibitors, Dynasore and recombinant PCSK9 protein, as well as by an LDL uptake promoter, Simvastatin. Taken together, this method provides a medium-to-high throughput platform for simultaneously screening pharmacological activity as well as monitoring of cell morphology, hence cytotoxicity of compounds regulating LDL influx. The analysis can be used with different imaging systems and analytical software.

Original languageEnglish (US)
Article numbere58564
JournalJournal of Visualized Experiments
Volume2018
Issue number141
DOIs
StatePublished - Nov 1 2018

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Cholesterol
LDL Cholesterol
Assays
Health
Imaging techniques
Monitoring
Recombinant proteins
Endothelial cells
Pathology
Cytotoxicity
Imaging systems
Screening
Cells
Throughput
Simvastatin
Kidney Diseases
Endocytosis
Recombinant Proteins
Coronary Vessels
Cardiovascular Diseases

Keywords

  • Atherosclerosis
  • Biology
  • Cholesterol
  • Dynamin
  • Dynasore
  • Issue 141
  • LDL influx
  • LDL uptake
  • LDLR
  • Lipids
  • Low density lipoprotein
  • Recombinant PCSK9
  • Simvastatin

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

LDL cholesterol uptake assay using live cell imaging analysis with cell health monitoring. / Ritter, Portia; Yousefi, Keyvan; Ramirez, Juliana; Dykxhoorn, Derek M; Mendez, Armando J; Shehadeh, Lina.

In: Journal of Visualized Experiments, Vol. 2018, No. 141, e58564, 01.11.2018.

Research output: Contribution to journalArticle

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