Quorum Sensing

(sharon) #1

3 Methods


3.1 Transepithelial
Electrical Resistance
(TER) and Paracellular
Flow



  1. TER and paracellular flux is evaluated using a two-
    compartment cell culture model.

  2. Caco-2 cells are seeded at a density of 10^5 cells/cm^2 onto
    Costar Transwell permeable culture inserts (pore size 3μm,
    Corning Inc., Corning, NY, USA) in standard 24-well plates.

  3. Cells are cultivated in GM at 37Cin5%CO 2 and maintained
    for 10–12 days until a tight monolayer is formed and a stable
    TER is attained. TER of the cell monolayers is measured using
    an epithelial volt-ohm meter with a stick-type electrode as per
    the manufacturer’s instructions (Fig.1). Only cell monolayers
    with a stable TER of 500–700Ωcm^2 and above should be
    used.

  4. TER value is calculated via the following equation: TER
    [Ωcm^2 ]¼(measured resistance for membrane with cells
    [Ω]measured black resistance for membrane without cells


Fig. 1Transepithelial electrical resistance (TER) setup. Caco-2 cell monolayers
are grown in 24-well plates on Costar Transwell permeable culture inserts (3μm
pore size). TER resistance of Caco-2 cell monolayers is measured using an
epithelial volt-ohm meter with stick-type electrodes (E1 and E2). TER
[Ωcm^2 ]¼(measured resistance for membrane with cells [Ω]measured
blackresistance for membrane without cells [Ω])area of the membrane
(0.33 cm^2 )

Autoinducer Effects on Mammalian Cells 217
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