The polarity of choroid plexus epithelial cells in vitro is improved in serum-free medium.
The influence of culture conditions on the development of normal characteristics of the choroid plexus epithelium has been investigated in vitro with respect to polarity, barrier properties, transport, and secretory activity. Withdrawal of serum supplement in the culture medium of cells grown on filters caused morphologically visible changes by an increased trimming of microvilli at the apical membrane side, which is accompanied by an increased expression of the Na+,K+-ATPase. Moreover cells under serum-free conditions exhibit structural changes in tight junctional zonula occludens protein-1 (ZO-1) organization, a reduced permeability, and a drastically increased electrical resistance from 150 ohms x cm2 in the presence of serum to 1,500 ohms x cm2 after serum withdrawal. Under these conditions, cell monolayers are able to build up a transcellular proton gradient and to secrete fluid into the upper (apical) filter compartment, which is accompanied by a polarized secretion of proteins like transthyretin. Active transport of the dyes fluorescein and phenol red by the organic anion transporter is found to be driven by the Na+,K+-ATPase. We come to the conclusion that removal of serum favors the differentiation process of the plexus epithelium in vitro, which brings the cell culture model closer to the physiological situation in vivo. We present preliminary evidence that epidermal growth factor may be one component in serum preventing the proper in vitro differentiation.
Institut für Biochemie, Westfälische Wilhelms-Universität Münster, Germany.
A
A
Hakvoort
M
M
Haselbach
J
J
Wegener
D
D
Hoheisel
H J
HJ
Galla
D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D001812:Blood-Brain Barrier / Q000502:physiology; D002417:Cattle; D016764:Cell Polarity / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D002831:Choroid Plexus / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D013552:Swine; D062826:Zonula Occludens-1 Protein
D016895:Culture Media, Serum-Free; D008565:Membrane Proteins; D010750:Phosphoproteins; D062826:Zonula Occludens-1 Protein