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A Fluorescence-based Assay for Characterization and Quantification of Lipid Droplet Formation in Human Intestinal Organoids
Published on: October 13, 2019
Human fetal intestinal epithelial cells metabolize and incorporate branched chain fatty acids in a structure specific
Lei Liu1, Zhen Wang2, Hui Gyu Park2
1College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan 410128, China; Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853, USA.
Insights
Branched chain fatty acids (BCFA) are incorporated into human fetal intestinal cells, demonstrating their metabolism and utilization. These findings suggest specific pathways in the fetal GI tract for handling BCFA, important for infant health.
Area of Science:
- Gastroenterology
- Cell Biology
- Biochemistry
Background:
- Branched chain fatty acids (BCFA) are present in the gastrointestinal (GI) tract of healthy newborn infants.
- BCFA have been shown to reduce necrotizing enterocolitis (NEC) in a neonatal rat model.
- BCFA are incorporated into the small intestine cellular lipids in vivo.
Purpose of the Study:
- To investigate the uptake, metabolism, and incorporation of BCFA into human fetal intestinal cells in vitro.
- To understand how human fetal intestinal cells handle and process BCFA.
Main Methods:
- Human H4 cells (fetal small intestine cell line) were incubated with specific albumin-bound non-esterified BCFA (anteiso-17:0, iso-16:0, iso-18:0, iso-20:0).
- Fatty acid (FA) profiles in cellular lipid fractions were analyzed post-incubation.
Main Results:
- All tested BCFA were readily incorporated as major constituents of cellular lipids.
- Anteiso-17:0 showed preferential uptake and was most effective at displacing normal fatty acids.
- Cholesterol esters (CE) demonstrated the highest affinity for BCFA incorporation (65%), while phospholipids (PL) incorporated 42% BCFA without cell death.
- BCFA underwent elongation and chain shortening, with iso-20:0 being chain shortened but not elongated.
Conclusions:
- Nontransformed human fetal intestinal epithelial cells efficiently incorporate and metabolize available BCFA.
- Metabolism of BCFA is structure-specific within these cells.
- The study implies the presence of specific pathways for handling BCFA in the human fetal GI tract, potentially influenced by vernix-derived BCFA during late gestation.
Background:
Branched chain fatty acids (BCFA) are constituents of gastrointestinal (GI) tract in healthy newborn human infants, reduce the incidence of necrotizing enterocolitis (NEC) in a neonatal rat model, and are incorporated into small intestine cellular lipids in vivo. We hypothesize that BCFA are taken up, metabolized and incorporated into human fetal cells in vitro.
Methods:
Human H4 cells, a fetal non-transformed primary small intestine cell line, were incubated with albumin-bound non-esterified anteiso-17:0, iso-16:0, iso-18:0 and/or iso-20:0, and FA profiles in lipid fractions were analyzed.
Results:
All BCFA were readily incorporated as major constituents of cellular lipids. Anteiso-17:0 was preferentially taken up, and was most effective among BCFA tested in displacing normal (n-) FA. The iso BCFA were preferred in reverse order of chain length, with iso-20:0 appearing at lowest level. BCFA incorporation in phospholipids (PL) followed the same order of preference, accumulating 42% of FA as BCFA with no overt morphological signs of cell death. Though cholesterol esters (CE) are at low cellular concentration among lipid classes examined, CE had the greatest affinity for BCFA, accumulating 65% of FA as BCFA. BCFA most effectively displaced lower saturated FA. Iso-16:0, iso-18:0 and anteiso-17:0 were both elongated and chain shortened by ±C2. Iso-20:0 was chain shortened to iso-18:0 and iso-16:0 but not elongated.
Conclusions:
Nontransformed human fetal intestinal epithelial cells incorporate high levels of BCFA when they are available and metabolize them in a structure specific manner. These findings imply that specific pathways for handling BCFA are present in the lumen-facing cells of the human fetal GI tract that is exposed to vernix-derived BCFA in late gestation.
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