Electrical resistance and macromolecular permeability of brain endothelial monolayer cultures.
Electrophysiological measurements were made on endothelial cells initially isolated as individual clones from bovine brain microvessels, and then grown as monolayers on a permeable support of glutaraldehyde-treated collagen gel. When transendothelial cell resistance (R) of the clones was measured, there was a range of values from a low of 157.4 +/- 4.5 omega.cm2 (n = 6) to a high of 783.2 +/- 7.0 omega.cm2 (n = 34). With the high-resistance cells, there was also a small potential difference of -0.46 +/- 0.03 mV luminal-side negative (n = 34). In comparison, endothelial cells from bovine aortas and rat epididymal fat pads cultured on the collagen gels had transendothelial R values of 13.5 +/- 0.2 (n = 62) and 0.45 +/- 0.03 (n = 10) omega.cm2, respectively. Exposure of the high-resistance brain endothelial cell monolayers to a Ca2+-free medium for 10 min decreased the R to 75% of the control values. Addition of Ca2+ back to the medium caused a return of the transendothelial R to control values within 1 h. Endothelial cells were also grown to confluency on microcarrier beads for permeability measurements to Evans blue dye-bovine serum albumin. Microcarriers with no cells (control) and microcarriers with bovine and epididymal endothelial cell monolayers showed no difference in the amount of adsorbed dye. Microcarriers with brain endothelial monolayers excluded up to 80% of the dye. This mammalian brain endothelial culture system will be a useful model for studies of the electrophysiological and permeability properties of the blood-brain barrier.
Department of Anatomy and Cellular Biology, Harvard Medical School, Boston, MA 02115.
M J
MJ
Rutten
R L
RL
Hoover
M J
MJ
Karnovsky
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002199:Capillary Permeability / Q000187:drug effects; D002417:Cattle; D002478:Cells, Cultured; D002560:Cerebrovascular Circulation; D004553:Electric Conductivity; D004594:Electrophysiology; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D004822:Epididymis / Q000166:cytology / Q000378:metabolism; D046911:Macromolecular Substances / Q000378:metabolism; D008297:Male; D051381:Rats
D046911:Macromolecular Substances; D002118:Calcium