Hydrocortisone reinforces the blood-brain properties in a serum free cell culture system.
The increasing number of newly developed drugs demands for functional in vitro models of the blood-brain barrier to determine their brain uptake. Cultured cerebral capillary endothelial cells are considered to be such a model, however in serum containing media they exhibit low electrical resistances and high permeabilities compared to the in vivo situation. Here we report the establishment of a serum-free cell culture model. Withdrawal of serum already caused a twofold increase of transendothelial resistance (TER), which in presence of serum is about 100-150 Omega x cm2. We tested several supplements and found that hydrocortisone is a potent stimulator for the formation of barrier properties. TERs up to 1000 Omega x cm2 were measured in the presence of physiological relevant hydrocortisone concentrations. In correspondence to the TER increase hydrocortisone decreased cell monolayer permeability for sucrose down to 5x10(-7) cm/s, which is close to the in vivo value of 1.2x10(-7) cm/s and by a factor of five lower compared to cultures without hydrocortisone and in presence of serum.
Institut für Biochemie, Westfälische Wilhelms-Universität, Münster, Germany.
D
D
Hoheisel
T
T
Nitz
H
H
Franke
J
J
Wegener
A
A
Hakvoort
T
T
Tilling
H J
HJ
Galla
D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects* / Q000502:physiology; D002196:Capillaries / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D002199:Capillary Permeability / Q000187:drug effects; D002200:Capillary Resistance / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002540:Cerebral Cortex / Q000098:blood supply / Q000166:cytology; D016895:Culture Media, Serum-Free; D004353:Drug Evaluation, Preclinical; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006854:Hydrocortisone / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology; D008954:Models, Biological; D013395:Sucrose / Q000493:pharmacokinetics; D013552:Swine
D016895:Culture Media, Serum-Free; D007328:Insulin; D013395:Sucrose; D004815:Epidermal Growth Factor; D006854:Hydrocortisone