Functional characterization of three clones of the human intestinal Caco-2 cell line for dietary lipid processing.
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 (Chantret et al. 1994). 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.
Salvini
Séverine
S
Unité 476 INSERM, Nutrition humaine et lipides, Marseille, France.
Charbonnier
Monique
M
Defoort
Catherine
C
Alquier
Christian
C
Lairon
Denis
D
D000704:Analysis of Variance; D053283:Apolipoprotein B-48; D001055:Apolipoproteins B / Q000378:metabolism; D018938:Caco-2 Cells / Q000378:metabolism*; D002791:Cholesterol, Dietary / Q000378:metabolism; D002999:Clone Cells / Q000378:metabolism; D004041:Dietary Fats / Q000378:metabolism*; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D007408:Intestinal Absorption; D019301:Oleic Acid / Q000378:metabolism
D053283:Apolipoprotein B-48; D001055:Apolipoproteins B; D002791:Cholesterol, Dietary; D004041:Dietary Fats; D019301:Oleic Acid; D006903:Hydroxymethylglutaryl CoA Reductases