Immortalized Human Endoneurial Endothelial Cells (THEndEC)

As part of the blood-nerve barrier (BNB), the endoneurial endothelial plays a significant role in the regulation of influx and efflux of water, nutrients and xenobiotics that is needed to maintain the peripheral nerve internal microenviroment.
The Immortalized Human Endoneurial Endothelial Cells (THEndEC) is a useful model for use in facilitated receptor-mediated in vitro BNB permeability and/or kinetic studies, in addition to susceptibility to drug intoxication research as it retained specialized transporters (i.e. P-gp, γ-GT, LAT-1, CRT and MCT-1) and restrictive barrier function similar to primary endoneurial endothelial cells. The THEndEC has been characterized to show specific binding to UEA-1 lectin and the ability of Dil-Ac-LDL uptake, indicative of α-L-fucose expression and specialized scavenger receptor expression, respectively. Further studies demonstrated transendothelial electrical resistance (TEER) and permeability to fluoresceinated sodium and dextran using THEndEC, making this cell line suitable for peripheral nerve permeability studies as well.

Immortalization Method: Serial passaging and stable transfection with a plamid carrying SV40 Large T antigen

BioSafety Level: II

Species: Human

Source Organ: Sciatic nerve

Growth Properties: Adherent

Morphology: Spindle shaped

Population Doubling: Variable doubling time from 28 to 53 hours

Seeding Density: 20,000-40,000 cells/cm²

Markers: GLUT-1, P-gp, γ-GT, LAT-1, CRT and MCT-1

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow II medium available at abm (TM002). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, 1 ng/ml recombinant human FGF2 (Z101455), 10 U/ml heparin, 50 µg/ml endothelial cell growth supplement, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Immunocytochemistry was used to confirm the nuclear expression of SV40-LTA in the immortalized cells; 2) RT-PCR and western blot were used to assess the expression of CRT, GLUT-1, P-gp, LAT-1, MCT-1 and γ-GT; 3) Transendothelial electrical resistance (TEER) assessment was performed to evaluate the biophysical property of the immortalized cells; 4) Sodium fluorescein and FITC-dextran were used to evaluate the solute permeability of the immortalized cells; 5) 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine (DiI)-labelled Ac-LDL was used to evaluate Ac-LDL uptake.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C