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Development of a relevant cell culture model to examine lutein absorption in vitro (645.12)
Author(s) -
Kamil Alison,
Blumberg Jeffrey,
Chen CY.
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.645.12
Subject(s) - lutein , micelle , caco 2 , chemistry , absorption (acoustics) , bioavailability , carotenoid , small intestine , intestinal epithelium , cell culture , in vitro , chromatography , biochemistry , food science , biology , pharmacology , epithelium , organic chemistry , materials science , genetics , aqueous solution , composite material
Lutein bioavailability is influenced by dietary fat which facilitates absorption via micelles. Permeable support systems are employed in in vitro nutrient absorption studies but data are absent on whether this approach generates results more relevant to in vivo absorption than a monolayer system (MONO). We used Caco‐2 colorectal cells to compare the efficacy of lutein uptake in a MONO and a permeable support Transwell ® system (TRAN) to examine the role of micelles. After plating for 2 and 21 d to attain confluence and differentiation in MONO and TRAN, respectively, cells were treated with 4 µg/mL lutein in 0.8% ethanol or synthetic micelles. After 36 h, lutein in cell lysate and basolateral medium was quantified by HPLC. Lutein in the cell lysate after ethanol was 220% greater than after micelle solubilization in MONO (3.0 vs. 1.4 µg/mg protein) but in the TRAN, lutein after ethanol was 77% lower (0.3 vs. 1.3). In TRAN, <1% (14.6 ng) of the micellized lutein was secreted to the basolateral medium while none was detected with lutein in ethanol. Thus, differentiation and/or culture system mediate lutein uptake into Caco‐2 and micelle components facilitate the intestinal secretion of lutein. As differentiated Caco‐2 display small intestinal phenotypes, micelles appear necessary to enable lutein transport across the small intestine but lutein can move into the large intestine efficiently without this facilitation. Grant Funding Source : Suported by USDA