Porcine Choroid plexus epithelial cells in culture: regulation of barrier properties and transport processes.
The epithelial cells of the choroid plexus are the structural basis of the blood-cerebrospinal fluid (CSF)-barrier. Here we summarise our recent efforts to culture those cells mainly on permeable supports in vitro. Isolated from porcine brains, we report a simple protocol for the primary culture using cytosine arabinoside as an additive that is cytotoxic for other cells except the plexus epithelial cells. Enhanced barrier properties are obtained by withdrawal of serum from the culture medium after confluency is reached. Cells improve their polarity, permeability for hydrophilic substrates is lowered, electrical resistance is increased tenfold, and a pH-gradient is built up across the cell monolayer. Polarised secretion of proteins and most importantly fluid secretion into the apical filter compartment was attained and proven to be dependent on the Na(+),K(+)-ATPase activity. Active transport processes (penicillin G, riboflavin, myo-inositol, ascorbic acid) were studied and clearly showed the involvement of the organic anion transporter. The permeability of the barrier was found to be regulated by cyclic adenosine monophosphate (cAMP). Moreover, we report that cell proliferation and differentiation is controlled by components of the extracellular matrix. The present culture model could now be used as an in vitro system to quantify drug transport across the blood-CSF-barrier.
Haselbach
M
M
Institut für Biochemie, Westfälische Wilhelms-Universität, D-48149 Münster, Germany.
Wegener
J
J
Decker
S
S
Engelbertz
C
C
Galla
H J
HJ
D000818:Animals; D001693:Biological Transport, Active; D002478:Cells, Cultured; D002555:Cerebrospinal Fluid / Q000378:metabolism; D002831:Choroid Plexus / Q000166:cytology* / Q000378:metabolism; D016895:Culture Media, Serum-Free; D000242:Cyclic AMP / Q000502:physiology; D004847:Epithelial Cells / Q000502:physiology; D005109:Extracellular Matrix / Q000502:physiology; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013552:Swine
D016895:Culture Media, Serum-Free; D000242:Cyclic AMP; D000254:Sodium-Potassium-Exchanging ATPase