The blood-brain barrier protects the central nervous system from potentially harmful substances by controlling the passage of cells and molecules. This barrier consists of vascular tissue, without pores, where the endothelial cells form tight junctions that allow only for select molecules to pass via pinocytosis. Immortalized Rat Brain Vascular Endothelial Cells (GPNT) was derived from microvessels in the cerebral cortex. The cells were immortalized by lipofection using an SV40 retroviral vector with a temperature sensitive large T antigen (tsA58) and contain a neomycin resistance gene. This cell line retains many characteristics of its primary parent cell line including expression of central nervous system specific markers, such as P-glycoprotein and GLUT-1, a high affinity glucose transporter not found in other vascular endothelial tissue. In addition, this cell line also expresses important immunological molecules including VCAM-I, PECAM-1 and MHC class I and II. Due to these properties this cell line may be useful in investigating immunological function, signal transduction, and migration of immune cells in specialized brain vascular endothelial tissue. More generally this cell line is ideal for research on the blood brain barrier.
Immortalization Method: Immortalized with retroviral vector containing SV40
BioSafety Level: II
Species: Rat
Source Organ: Blood Vessel
Growth Properties: Adherent
Morphology: Epithelial
Population Doubling: N/A
Seeding Density: N/A
Markers: P-glycoprotein, GLUT-1, the transferrin receptor. Neomycin resistant. von Willebrand factor, RECA-1
Donor Age: N/A
Donor Gender: N/A
Donor Ethnicity: N/A
Propagation: Use of Type I Calf Skin Collagen (Sigma) is required for cell adhesion to the culture vessels. All plasticware must be pre-coated for several hours at 10 μg/cm² from a 0.01% solution in acetic acid. After removal the coated plasticware is exposed to ammonia vapour for 5 min. Plasticware must be rinsed thoroughly with PBS prior to use. Grow cells in coated culture vessels with the following conditions.The base medium for this cell line is Ham’s F10 Medium (Lonza). To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum (TM999) to a final concentration of 10%, L-glutamine (G275) to a final concentration of 2 mM, Recombinant Rat FGF2 (Z300005) to a final concentration of 2 ng/mL, and heparin to a final concentration of 80 μg/mL. Change media every 2-3 days. Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
Quality Control: N/A
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C