Primary Cultures of Brain Capillary Endothelial Cells

Franke et al. (1999) have described an in vitro model for screening of drug entry into the brain using primary cultures of porcine brain capillary endothelial cells (PBCEC). By using serum-free culture conditions 

Organic Solute Transporter Organic Anion Transporting ABC Transporter Family  

Names Expression Names Expression Names Expression 

Collagen fibers are collected, as described by Bomstein et al. (1958). Fibers of each tail are incubated in 50 ml 0.1 % (v/v) acetic acid at 4 °C for 24 h. Centrifugation for 2 h at 5000 x g separates the fiber residues from solubilized collagen which is aspirated. Total protein content of the collagen solution is determined and adjusted to 0.1 mg/ml. Transwell or Transwell-Clear filters are coated by adding 100 pi of rat-tail collagen solution per 

Directly prior the experiment the quality, that is the tightness, of the PBCEC monolayer is assessed by determination of the transendothelial electrical resistance (TEER) of each filter. TEER measurements can be easily performed with the ENDOHM-24 chamber and the EVOHM voltometer (World Precision Instruments, Inc,). The resistance data, expressed in Q, are multiplied with the filter surface (4.52 cm2), corrected for blank filter resistance and expressed in L2 x cm2. 

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The presence at the BBB of members of the multidrug resistance-associated protein (MRPs) family, whose members preferentially transport anionic compounds, is still controversial. The seven members of the MRP family belong, like P-gp, to the ATP-binding cassette (ABC) protein superfamily. Mrpl has been found at the BBB in isolated rat brain capillaries, primary cultures of brain capillary endothelial cells and in immortalized capillary endothelial cells, but not in human brain capillaries [59]. Another member, MRP2 has been found at the luminal membrane of the brain endothelial cells [60]. However, further studies are required to show that there are MRP transporters at the BBB (Figure 15.5). As for P-gp, a functional Mrpl was found in primary cultured rat astrocytes [56] and it has been shown to take part in the release of glutathione disulfide from brain astrocytes under oxidative stress [61]. 

II.1.1.1 Primary Cultures of Brain Capillary Endothelial Cells 522... 

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