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Cited by 32Lipid metabolism and biosynthesisCancer, Lipids, and MetabolismLipid metabolism and disorders

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Oleic acid differentially affects lipid droplet storage of de novo synthesized lipids in hepatocytes and adipocytes

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Castillo, Hannah B., Shuster, Sydney O., Tarekegn, Lydia H., Davis, Caitlin M.·2024·Chemical Communications
DOI 10.1039/d3cc04829bPMC PMC10939124

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Abstract

This work introduces a multiplexed sub-micron vibrational imaging technique to monitor differential effects of oleic acid in adipocytes and hepatocytes.

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Fig. 1 Schematic of experiments and data collection. (A) Isotope labelling of de novo lipogenesis and fatty-acid scavenging pathways where oleic acid is surmised to inhibit ACC. (B) Live cell data collection by OPTIR. Cells grown on a calcium fluoride cover slip are mounted in PBS using a spacer. (C) Multispectral imaging at single frequencies.
Fig. 1 Schematic of experiments and data collection. (A) Isotope labelling of de novo lipogenesis and fatty-acid scavenging pathways where oleic acid is surmised to inhibit ACC. (B) Live cell data collection by OPTIR. Cells grown on a calcium fluoride cover slip are mounted in PBS using a spacer. (C) Multispectral imaging at single frequencies.CC BY · PubMed Central
Fig. 2 Live cell characterization of the cell lines (A) Huh-7 and (B) differentiated 3T3-L1 after feeding with 2H oleic acid (60 μM) and 13C glucose (4.5 mg mL−1). Scale bars are 20 μm. (C) Full IR spectra of a point within a fixed Huh-7 cell LD 48 hours after feeding 2H OA. 2H-C from the 2H oleic acid (green, structure in inset) which has been esterified in LDs. 2214 cm−1 is assigned to 2H-C stretching. 13C <svg xmlns="http://www.w3.org/2000/svg" version="1.0" width="13.200000pt" height="16.000000pt" viewBox="0 0 13.200000 16.000000" preserveAspectRatio="xMidYMid meet"><metadata> Created by potrace 1.16, written by Peter Selinger 2001-2019 </metadata><g transform="translate(1.000000,15.000000) scale(0.017500,-0.017500)" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg> O stretch at 1703 cm−1 where the 13C comes from metabolized 13C glucose (blue, structure in inset). Pictured alongside the unlabelled 12CO band at 1744 cm−1.
Fig. 2 Live cell characterization of the cell lines (A) Huh-7 and (B) differentiated 3T3-L1 after feeding with 2H oleic acid (60 μM) and 13C glucose (4.5 mg mL−1). Scale bars are 20 μm. (C) Full IR spectra of a point within a fixed Huh-7 cell LD 48 hours after feeding 2H OA. 2H-C from the 2H oleic acid (green, structure in inset) which has been esterified in LDs. 2214 cm−1 is assigned to 2H-C stretching. 13C <svg xmlns="http://www.w3.org/2000/svg" version="1.0" width="13.200000pt" height="16.000000pt" viewBox="0 0 13.200000 16.000000" preserveAspectRatio="xMidYMid meet"><metadata> Created by potrace 1.16, written by Peter Selinger 2001-2019 </metadata><g transform="translate(1.000000,15.000000) scale(0.017500,-0.017500)" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg> O stretch at 1703 cm−1 where the 13C comes from metabolized 13C glucose (blue, structure in inset). Pictured alongside the unlabelled 12CO band at 1744 cm−1.CC BY · PubMed Central
Fig. 3 Visualization of rates of glucose-derived de novo lipid storage in live Huh-7 cells fed with both 2H OA (60 μM) and 13C glucose (4.5 mg mL−1). (A) Brightfield images of live cells. (B) 2H OA ratio images of 2H lipid to the 12C lipid band, after correction. (C) 13C glucose ratio images of the 13C lipid band to the 12C lipid band, after correction. (D) Representative spectra of the live Huh-7 cell at the 48-hour time point both inside a LD (pink) and outside the droplet (blue) (E) Average ratios of 2H lipid band (grey) and 13C lipid band (red) to 12C lipid band across many cells (N = 14). Scale bars are 10 μm.
Fig. 3 Visualization of rates of glucose-derived de novo lipid storage in live Huh-7 cells fed with both 2H OA (60 μM) and 13C glucose (4.5 mg mL−1). (A) Brightfield images of live cells. (B) 2H OA ratio images of 2H lipid to the 12C lipid band, after correction. (C) 13C glucose ratio images of the 13C lipid band to the 12C lipid band, after correction. (D) Representative spectra of the live Huh-7 cell at the 48-hour time point both inside a LD (pink) and outside the droplet (blue) (E) Average ratios of 2H lipid band (grey) and 13C lipid band (red) to 12C lipid band across many cells (N = 14). Scale bars are 10 μm.CC BY · PubMed Central
Fig. 4 Effect of OA feeding on rates of de novo lipid storage. Ratio of 13C lipid to 12C lipid in live (A) Huh-7 and (B) 3T3-L1 cells fed with both 2H oleic acid and 13C glucose (red) or BSA vehicle and 13C glucose (purple) at 24, 48, and 72 h. N = 8–16 cells per time point *p-value < 0.05 **p-value < 0.005 ***p-value < 0.0005 N.S. p-value > 0.05.
Fig. 4 Effect of OA feeding on rates of de novo lipid storage. Ratio of 13C lipid to 12C lipid in live (A) Huh-7 and (B) 3T3-L1 cells fed with both 2H oleic acid and 13C glucose (red) or BSA vehicle and 13C glucose (purple) at 24, 48, and 72 h. N = 8–16 cells per time point *p-value < 0.05 **p-value < 0.005 ***p-value < 0.0005 N.S. p-value > 0.05.CC BY · PubMed Central

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Metadata

Authors
Castillo, H. B., Shuster, S. O., Tarekegn, L. H., & Davis, C. M.
Affiliations
Yale University (US)
Corresponding author
Davis, Caitlin M.
ORCID
Venue
Chemical Communications
Publisher
Royal Society of Chemistry (RSC)
Published
2024
Volume
60
Issue
23
Pages
3138-3141
ISSN
  • 1359-7345 (ISSN-L)
  • 1364-548X
DOI
10.1039/d3cc04829b
PMC
PMC10939124
OpenAlex
W4391347979
Cited by
32
References
42
Citations by year
2026: 16 · 2025: 8 · 2024: 8
FWCI
5.14
Type
journal article
Language
en
Open access
Licence
Locations
  • Chemical Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by-nc-nd)
Funders
  • National Science Foundation (DGE-2139841)
  • Research Corporation for Science Advancement
  • Gordon and Betty Moore Foundation
  • National Institute of General Medical Sciences (R35 GM151146, T32 GM008283)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Lipid metabolism and disorders
Keywords
oleic acid, de novo lipogenesis, hepatocytes, adipocytes, lipid metabolism

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CoChat. (2026). Citation check: Oleic acid differentially affects lipid droplet storage of de novo synthesized lipids in hepatocytes and adipocytes. https://app.cochat.ai/papers/10.1039/d3cc04829b
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Castillo, H. B., Shuster, S. O., Tarekegn, L. H., & Davis, C. M. (2024). Oleic acid differentially affects lipid droplet storage of de novo synthesized lipids in hepatocytes and adipocytes. Chemical Communications. https://doi.org/10.1039/d3cc04829b