Development of an automated 7-day 96-well Caco-2 cell culture model.
Caco-2 cells are a widely accepted model to predict permeability and absorption of compounds in humans. We built up an automated 96-wellplate Caco-2 permeation model with reduced growth time. Model compounds (metoprolol, ketoprofen, verapamil, naproxen, hydrochlorthiazide), permeability markers (TEER, Lucifer Yellow, D-[1-(14)C]-mannitol) and confocal microscopy were used to assess the utility of our method on Biomek-FX-automation. The confocal imaging data showed that Caco-2 cells formed monolayers when cultured for 7 days with initial cell density of 2.1 x 10(5) cells/cm2. P-Glycoprotein was present in Caco-2 cells, localized on the plasma membrane. Permeation of model compounds was comparable to those obtained from traditional 12-wellplate experiments.
Galkin
A
A
Drug Discovery and Development Technology Center (DDTC) and Division of Pharmaceutical Biology, Faculty of Pharmacy, University of Helsinki, Finland.
Pakkanen
J
J
Vuorela
P
P
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000465:Algorithms; D001331:Automation; D015415:Biomarkers; D018938:Caco-2 Cells / Q000378:metabolism*; D002452:Cell Count; D002463:Cell Membrane Permeability / Q000187:drug effects* / Q000502:physiology; D002851:Chromatography, High Pressure Liquid; D017097:Electric Impedance; D005456:Fluorescent Dyes; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D007546:Isoquinolines; D018613:Microscopy, Confocal
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000199:Actins; D015415:Biomarkers; D005456:Fluorescent Dyes; D007546:Isoquinolines; C017475:lucifer yellow