abm | Immortalized Mouse Cerebral Capillary Endothelial Cells (cEND) | T0290

abm | Immortalized Mouse Cerebral Capillary Endothelial Cells (cEND) | T0290

in vivo, specifically in that 1) it retains the endothelial markers VE-Cadherin and PECAM-1; 2) it displays tight junction associated markers such as occludin, claudin-3, -5, -12, 3) it expresses the BBB marker protein Glut-1 and 4) it shows high electrical resistance, representing barrier properties. This cell line is also responsive to glucocorticoid, estrogen-treatment and pro-inflammatory mediator such as TNFα, making this cell line valuable in elucidating cellular responses of endothelial cells to different stimuli. Together with the Immortalized Mouse Cerebellar Capillary Endothelial Cell Line (cerebEND), the two cell lines represent important in vitro model system for these different brain regions.

Immortalization Method:

Transformation with oncoprotein of murine Polyomavirus, Polyoma middle T antigen (PymT)

BioSafety Level:

II

Organism:

Neonatal mouse (WT 129 Sv strain)

Species:

MOuse

Source Organ:

Cerebral cortex

Organ Type:

Brain

Growth Properties:

Adherent

Morphology:

Spindle shaped

Passage Number:

N/A

Population Doubling:

60-80 hours

Seeding Density:

30,000-35,000 cells/cm10²; Recommended split ratio: no greater than 1:4

Markers:

Claudin-5, Occuldin, Glut-1, VE-Cadherin

Donor Age:

N/A

Donor Gender:

N/A

Donor Ethnicity:

N/A

Propagation:

The base medium for this cell line is Prigrow III medium available at abm . To make the completed growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum to final concentration of 10% and Penicillin/Streptomycin Solution to a final concentration of 1%
Carbon dioxide (CO₂): 5%, Temperature: 37.0°C.

Quality Control:

1) Endothelial marker VE-Cadherin and PECAM-1 assessed by immunostainning; 2) Tight junction protein Claudin-5 assessed by immunostainning and claudin-1, -3 and -12 detected in mRNA level; 2) BBB marker protein Glut-1 and tight junction-associated protein occludin measured by western blot analysis. The cell line's response to pro-inflammatory stimuli was measured by 1) TER, 2) western blot and 3) gene expression analysis using RT-PCR.

Shipping Condition:

Dry Ice

Storage Condition:

liquid nitrogen or -180C