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Flow cytometric method for the detection of flavonoids in cell lines

Charlotte Grootaert (UGent) , Gerard Gonzales (UGent) , Hanne Vissenaekens (UGent) , Tom Van de Wiele (UGent) , Katleen Raes (UGent) , Guy Smagghe (UGent) and John Van Camp (UGent)
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Abstract
Here, we describe an easy-to-use flow cytometric method using diphenylboric acid 2-amino ethyl ester (DPBA) stain for the detection of flavonoids in cells from human/animal origin. Flavonoid bioavailability and bioactivity depend on structure, conjugation and the cell type to which they are presented. We have studied cellular uptake of five flavonoids with different structures and conjugation forms. First, parameters including fixation method, technical and batch variability, and concentration were optimized. Second, uptake of two aglyconesquercetin and hesperetinand their corresponding glycosidesrutin and hesperidinin Caco-2 cells was compared. Third, the aglycone quercetin, glycoside rutin, and glucuronide baicalin were added to the Caco-2, HepG2, and CHO-K1 cell lines at 1, 10, and 20 mu M concentrations and cellular uptake was measured after 1, 4, and 7 h. We conclude that quercetin was taken up by cells in a dose-dependent way, and that HepG2 cells had the highest uptake factors, followed by CHO-K1 and Caco-2 cells. Confocal microscopy showed cell type-dependent localization of quercetin in the cell membrane and cytoplasm. No uptake of flavonoid glycosides was detected. This flow cytometric method can be used for future research unravelling mechanisms behind flavonoid bioactivity in health and disease at the cellular level.
Keywords
flavonoid, cell lines, flow cytometry, DPBA, BETA-GLUCURONIDASE, CACO-2 CELLS, IN-VITRO, QUERCETIN, METABOLISM, MODEL, LOCALIZATION, PERMEABILITY, TRANSPORTERS, MRP2

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Chicago
Grootaert, Charlotte, Gerard Gonzales, Hanne Vissenaekens, Tom Van de Wiele, Katleen Raes, Guy Smagghe, and John Van Camp. 2016. “Flow Cytometric Method for the Detection of Flavonoids in Cell Lines.” Journal of Biomolecular Screening 21 (8): 858–865.
APA
Grootaert, C., Gonzales, G., Vissenaekens, H., Van de Wiele, T., Raes, K., Smagghe, G., & Van Camp, J. (2016). Flow cytometric method for the detection of flavonoids in cell lines. JOURNAL OF BIOMOLECULAR SCREENING, 21(8), 858–865.
Vancouver
1.
Grootaert C, Gonzales G, Vissenaekens H, Van de Wiele T, Raes K, Smagghe G, et al. Flow cytometric method for the detection of flavonoids in cell lines. JOURNAL OF BIOMOLECULAR SCREENING. 2016;21(8):858–65.
MLA
Grootaert, Charlotte, Gerard Gonzales, Hanne Vissenaekens, et al. “Flow Cytometric Method for the Detection of Flavonoids in Cell Lines.” JOURNAL OF BIOMOLECULAR SCREENING 21.8 (2016): 858–865. Print.
@article{7257165,
  abstract     = {Here, we describe an easy-to-use flow cytometric method using diphenylboric acid 2-amino ethyl ester (DPBA) stain for the detection of flavonoids in cells from human/animal origin. Flavonoid bioavailability and bioactivity depend on structure, conjugation and the cell type to which they are presented. We have studied cellular uptake of five flavonoids with different structures and conjugation forms. First, parameters including fixation method, technical and batch variability, and concentration were optimized. Second, uptake of two aglyconesquercetin and hesperetinand their corresponding glycosidesrutin and hesperidinin Caco-2 cells was compared. Third, the aglycone quercetin, glycoside rutin, and glucuronide baicalin were added to the Caco-2, HepG2, and CHO-K1 cell lines at 1, 10, and 20 mu M concentrations and cellular uptake was measured after 1, 4, and 7 h. We conclude that quercetin was taken up by cells in a dose-dependent way, and that HepG2 cells had the highest uptake factors, followed by CHO-K1 and Caco-2 cells. Confocal microscopy showed cell type-dependent localization of quercetin in the cell membrane and cytoplasm. No uptake of flavonoid glycosides was detected. This flow cytometric method can be used for future research unravelling mechanisms behind flavonoid bioactivity in health and disease at the cellular level.},
  author       = {Grootaert, Charlotte and Gonzales, Gerard and Vissenaekens, Hanne and Van de Wiele, Tom and Raes, Katleen and Smagghe, Guy and Van Camp, John},
  issn         = {1087-0571},
  journal      = {JOURNAL OF BIOMOLECULAR SCREENING},
  language     = {eng},
  number       = {8},
  pages        = {858--865},
  title        = {Flow cytometric method for the detection of flavonoids in cell lines},
  url          = {http://dx.doi.org/10.1177/1087057116653220},
  volume       = {21},
  year         = {2016},
}

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