Immortalized Rat Brain Vascular Endothelial Cells (GPNT)

Mfr.No.	T0685
Description	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.
Cell Type	Immortalized Cells
Biosafety Level	2
Depositor	UCLB
Disclaimer	1. 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 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. 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). 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. 5. 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."
Donor History	Lewis rat
Expression Region	P-glycoprotein, GLUT-1, the transferrin receptor, von Willebrand factor, RECA-1, puromycin-resistance, VCAM-I, PECAM-1, MHC class I, MHC class II
Growth Properties	Adherent, epithelial
Growth Conditions	PriCoat?ECM T25 Flasks (G999) or Applied Cell Extracellular Matrix (G422) are?equired?or cell adhesion to the culture vessels.?am? F10 Medium (Lonza) + heat-inactivated 10% FBS + 2 mM L-Glutamine (G275) + 2 ng/ml rrFGF2 (Z300005) + 80 ?g/ml Heparin + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.
Quantity	1x106 cells / 1.0 ml
Tissue	Blood Vessel
Morphology	Polygonal
Notes	1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use.To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Rat (R. norvegicus)
Population Doubling	48 - 60 hours
Species	Rat (R. norvegicus)
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: fetal bovine serum (TM999)* to a final concentration of 15% 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.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control	Western blot was used to quantify SV40 gene expression in immortalized cell line.

Applications Range	Research Use Only.

Shipping	Ship with dry ice.
Product Format	Frozen
Product Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol	Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10900893	T0685	1x10^6 cells / 1.0 ml	-		Add to Quote

Immortalized Rat Brain Vascular Endothelial Cells (GPNT)

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