Homeostasis of the central nervous system (CNS) is maintained by the blood brain barrier (BBB) and disruptions to the BBB are linked to many disorders of the CNS. The Immortalized Mouse Cerebral Capillary Endothelial Cell Line (cEND) provides a useful model for studies involving the differentiation and regulation of BBB as it shares principal features of the BBB 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 TNFalpha, 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.
Cell Type
Immortalized Cells
Disclaimer
1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item. 2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 mug, Cat.# C207, $450.00) or cell lysate (100 mug, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 5. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 6. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Expression Level
Claudin-5, Occuldin, Glut-1, VE-Cadherin
Growth Properties
Adherent
Quantity
1x106 cells / 1.0 ml
Tissue
Brain
Morphology
Spindle shaped
Population Doubling
60 - 80 hours
Species
Mouse (M. musculus)
Propagation
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum to a final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.The Differentiation Medium is composed of 2% heat-inactivated fetal bovine serum instead of 10%. Culturing cEND cells in serum-reduced medium leads to an increase in TER (from 150 omegacm2 in the presence of 10% serum to 500 omegacm2 in the presence of 2% serum), as well as change in cell morphology (from spindle-shaped to cobble-stone like). TER can be further increased by the addition of 110nM hydrocortisone (800 omegacm2 ) or 1µM insulin (1000 omegacm2 ) into the differentiation medium.
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.
Reviews of Immortalized Mouse Cerebellum Cells (CRBL)