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Print Version |
| Biosesafety Level |
II |
| Organism |
Homo sapiens |
| Source Organ |
Sciatic nerve |
| Growth Properties |
Adherent |
| Morphology |
spindle shaped |
| Population Doubling |
Variable doubling time from 28 to 53 hours |
| Recommended Seeding Density |
10,000-15,000 cells/cm2 |
| Markers |
GLUT-1, P-gp, γ-GT, LAT-1, CRT and MCT-1 |
| Applications |
For Research Use Only |
| Immortalization Method |
Serial passaging and stable transfection with a plamid carrying SV40 Large T antigen |
| Description |
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.
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| Procedure Overview |
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| Propagation |
Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow II medium available from ABM (TM002). To make the completed growth medium, add the following components to the base medium: 10% fetal bovine serum (TM999), 1 ng/ml recombinant human FGF2 (Z101455), 10 U/ml heparin, 50 µg/ml endothelial cell growth supplement and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C. |
| Preservation |
1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.2. Storage Temperature: Liquid nitrogen vapour phase. |
| 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 evalsuate the biophysical property of the immortalized cells; 4) Sodium fluorescein and FITC-dextran were used to evalsuate the solute permeability of the immortalized cells; 5) 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine (DiI)-labelled Ac-LDL was used to evalsuate Ac-LDL uptake. |
| Disclaimer |
1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. 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 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
2. We strongly 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 evalsuate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical locations. 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).
3. 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.
4. 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. |
