Functional characterization of three clones of the human intestinal Caco-2 cell line for dietary lipid processing

Abstract

We aimed to improve the use of the human intestinal Caco-2 cell line for studying dietary lipid and cholesterol processing by using isolated pure clones (). Three clones (TC7, PD7 and PF11) were grown as monolayers on semi-permeable filters and compared for cell viability, fatty acid and cholesterol apical uptake or basolateral secretion, apolipoprotein B-48 basolateral secretion and 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase activity. The TC7 clone showed the best viability upon apical incubation with mixed micelles and should be preferred for routine work. Short-term (3·0 h) rates of apical uptake of cholesterol were not different with the three clones, whereas the rate of apical uptake of oleic acid (18 : 1) was lower (P<0·05) with PF11 (250·6 nmol/mg) and the basolateral secretion of cholesterol and oleic acid was lower with the TC7 clone (0·40 and 29·1 nmol/mg respectively). The secretion of apolipoprotein B-48 basolaterally was about 2-fold lower than from PD7 clone. The basal levels of HMG-CoA reductase activity were significantly different (P<0·05; TC7>PF11>PD7). The down-regulation of the enzyme activity was moderate (range 13·8–21·0 %) and comparable in the presence of apical micellar cholesterol, but was much marked upon basolateral incubation with LDL (range 34·0–53·6 %), especially for the PD7 clone. In conclusion, the Caco-2 clones characterized here proved to be particularly suitable for studying lipid nutrients processing. Because these three clones exhibit some different metabolic capabilities, they provide a new tool to study intestinal response to lipid nutrients.

Fatty acids affect micellar properties and modulate vitamin D uptake and basolateral efflux in Caco-2 cells

“…Caco-2 clone TC-7 cells [19,20] were a gift from Dr. M. Rousset (UMR_S 872, Villejuif, France). Cells were cultured in the presence of DMEM supplemented with 16% heat-inactivated FBS, 1% non-essential amino acids and 1% antibiotics (complete medium), as previously described [15,21].…”

NPC1L1 and SR‐BI are Involved in Intestinal Cholesterol Absorption from Small‐Size Lipid Donors

“…Because of the heterogeneity of the parental population of Caco-2 cells, the selected experimental model was the TC7 clone of the human intestinal cell-line, which had previously been characterized for cholesterol absorption studies [29]. When the TCA concentration was increased (from 2 to 8 mM; Fig.…”

“…The TC7 cells, a clone of human intestinal cell-line Caco-2, were used between passages 30 and 50 as previously described [28,29]. Stocks of cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 25 mM glucose, supplemented with 1% nonessential amino acids and 20% heat-inactivated (30 min, 56°C) fetal calf serum in a 10% CO 2 humidified incubator at 37°C.…”

Vitamin E decreases endogenous cholesterol synthesis and apo-AI-mediated cholesterol secretion in Caco-2 cells

“…Caco-2 clone TC-7 cells [22,23] were a gift from Dr. M. Rousset (U178 INSERM, Villejuif, France). Cells were cultured in the presence of DMEM supplemented with 20% heat-inactivated FBS, 1% nonessential amino acids and 1% antibiotics (complete medium).…”