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bEnd.3 [BEND3]

CRL-2299

Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Cell type
endothelial cell
Morphology
endothelial
Tissue
Brain; Cerebral cortex
Disease
Endothelioma
Applications
3D cell culture
Neuroscience
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Price: $495.00 EA
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Documentation

Biosafety Icon BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications
The cells were transformed by infection with the NTKmT retrovirus vector that expresses polyomavirus middle T antigen.
The endothelial nature of these cells was confirmed by the observed expression of von Willebrand factor and uptake of fluorescently labeled low density lipoprotein (LDL).

Characteristics

Growth properties
Adherent
Derivation
The cells were transformed by infection with the NTKmT retrovirus vector that expresses polyomavirus middle T antigen.
Age
6 weeks
Strain
BALB/c
Immortalization method
Polyoma middle T antigen
Antigen expression
ICAM-1 +; VCAM-1 +; MAdCAM-1 +
Genes expressed
von Willebrand factor-; ICAM-1+; VCAM-1+; MAdCAM-1+ ; The expression of Peyer's Patch high endothelial receptor for lymphocytes, the mucosal vascular addressin (MAdCAM-1) and E-selectin can be induced on bEnd.
Comments
The endothelial nature of these cells was confirmed by the observed expression of von Willebrand factor and uptake of fluorescently labeled low density lipoprotein (LDL).
The expression of Peyer's Patch high endothelial receptor for lymphocytes, the mucosal vascular addressin (MAdCAM-1) and E-selectin can be induced on bEnd.3 cells by cytokines and lipopolysaccharide (LPS).
This induction by Tumor Necrosis Factor alpha (TNF alpha), interleukin 1 (IL-1) or LPS is concentration and time dependent.
MAdCAM-1 is expressed on the surface of unstimulated bEnd.3 cells at early passages but not at passages greater than 30.
Intracellular adhesion molecule 1 (ICAM-1) is constitutively expressed on the cells, and expression is increased by treatment with LPS, IL-1 and TNF alpha.
Vascular cell adhesion molecule 1 (VCAM-1) is constitutively expressed on the cells at early passages but not at passages over 30.
P-selectin can be induced on bEnd.3 cells by Tumor Necrosis Factor alpha (TNF alpha) at both early and late passages but expression is greater at passages over 30.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

         

Subculturing procedure
Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.03% (w/v) EDTA solution to remove all traces of serum which contains trypsin inhibitor.Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer just begins to detach. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended
Medium Renewal: Every 2 to 3 days
Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected

History

Depositors
EC Butcher
Special collection
NCRR Contract

Legal disclaimers

Intended 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.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

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. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

This material was deposited by by Dr. Werner Risau, The Max Planck Society and is distributed for research purposes only under the ATCC Material Transfer Agreement. This material has been released subject to the following:

  1. Transfers - the material or its products must not be transferred to third parties.
  2. Commercial Use - all for-profit and non-profit Recipients must obtain a commercial use license prior to Commercial Use.
  3. Publishing - in all papers reporting any use of these cells, or derived products, a direct reference will be made to the original publication (Montesano, et al. Cell 62:435-445, 1990).

Any proposed commercial use of these cells must first be negotiated with:

Max-Planck-Institut fur physiologische und klinische Forschung
W.G. Kerckoff Institut
Abteilung molekulare Zellbiologie
Parkstrasse 1, 61231 Bad Nauheim, Germany
Phone: 49-6032-705203/265
Website: https://www.mpi-hlr.de/contact-and-locations

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Montesano R, et al. Increased proteolytic activity is responsible for the aberrant morphogenetic behavior of endothelial cells expressing the middle T oncogene. Cell 62: 435-445, 1990. PubMed: 2379237

Sikorski EE, et al. The Peyer's patch high endothelial receptor for lymphocytes, the mucosal vascular addressin, is induced on a murine endothelial cell line by tumor necrosis factor-alpha and IL-1. J. Immunol. 151: 5239-5250, 1993. PubMed: 7693807

Williams RL, et al. Embryonic lethalities and endothelial tumors in chimeric mice expressing polyoma virus middle T oncogene. Cell 52: 121-131, 1988. PubMed: 3345558

Frequently Asked Questions

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Telephone

Telephone

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800-638-6597

Outside the US

+1-703-365-2700
hours

Hours of Operation

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