Quantification of Endothelial Fatty Acid Uptake using Fluorescent Fatty Acid Analogs

Area of Science:

• Cell Biology
• Metabolic Science
• Biochemistry

Background:

• Endothelial cells (ECs) are crucial for regulating fatty acid (FA) transport to metabolic tissues.
• Limited tools exist for reliable and scalable quantification of EC FA uptake.

Purpose of the Study:

• To develop a rapid, quantitative, and cost-effective assay for measuring FA uptake in ECs.
• To investigate chain-length-specific FA uptake dynamics.
• To provide a versatile platform for studying endothelial lipid transport.

Main Methods:

• Utilized fluorescent FA analogs (BODIPY-C12, BODIPY-C16) in a 96-well plate format.
• Incorporated positive (3-hydroxyisobutyrate, lactate) and negative (niclosamide) controls.
• Validated in primary (HUVECs) and immortalized (EA.hy926) EC lines, normalizing data with Hoechst staining.

Main Results:

• The assay accurately detects time- and dose-dependent FA uptake.
• Demonstrated chain length-specific uptake dynamics.
• Validated across different EC types and experimental conditions, including live-cell imaging.

Conclusions:

• This novel assay offers improved safety, speed, and versatility compared to traditional methods.
• Provides a robust platform for studying EC lipid transport regulation.
• Facilitates research into the role of endothelial FA uptake in metabolic diseases.