bEnd.3 ATCC ® CRL-2299™ Mus musculus brain/cerebral cortex

Permits	These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Organism	Mus musculus, mouse
Tissue	brain/cerebral cortex
Cell Type	endothelialpolyoma middle T antigen transformed
Product Format	frozen
Morphology	endothelial
Culture Properties	adherent
Biosafety Level	1
Disease	endothelioma
Age	6 weeks
Strain	BALB/c
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).
Storage Conditions	liquid nitrogen vapor temperature

Derivation	The cells were transformed by infection with the NTKmT retrovirus vector that expresses polyomavirus middle T antigen.
Antigen Expression	ICAM-1 +; VCAM-1 +; MAdCAM-1 +
Genes Expressed	von Willebrand factor,ICAM-1 +; VCAM-1 +; MAdCAM-1 +,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.
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.

Complete Growth 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%.
Subculturing	Protocol: 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. Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation	Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature
Culture Conditions	Temperature: 37.0°C Temperature: 37.0°C Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Name of Depositor	EC Butcher
Passage History	MAdCAM-1 is expressed on the surface of unstimulated bEnd.3 cells at early passages but not at passages greater than 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. Vascular cell adhesion molecule 1 (VCAM-1) is constitutively expressed on the cells at early passages but not at passages over 30.
References	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

Name of Depositor

EC Butcher

Passage History

MAdCAM-1 is expressed on the surface of unstimulated bEnd.3 cells at early passages but not at passages greater than 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.

Vascular cell adhesion molecule 1 (VCAM-1) is constitutively expressed on the cells at early passages but not at passages over 30.

References

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

Permits	These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation	Product Sheet Certificate of Analysis MSDS
Restrictions	The bEnd.3 cell line is provided for research use only under the following terms:1. bEnd.3 cells, or products derived from them, must not be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purpose of sale, or producing for sale, bEnd.3 cells or their products. Commercial interests are the exclusive property of The Max Planck Society. 2. Any proposed commercial use of bEnd.3 cells, or products derived from them, must first be negotiated with Dr. Britta Engelhardt, on behalf of the late Dr. Werner Risau, Max-Planck-Institut fur physiologische und klinische Forschung, W.G. Kerckoff Institut, Abteilung molekulare Zellbiologie, Parkstrasse 1, 61231 Bad Nauheim, Germany. Telephone: 49-6032-705203/265; FAX: 49-6032-072259; email: b.engelhardt@kerckoff.mpg.de3. In all papers reporting any use of bEnd.3 cells, or products derived from them, a direct reference will be made to the original publication (Montesano, et al. Cell 62:435-445, 1990).
References	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

bEnd.3 (ATCC® CRL-2299™)

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bEnd.3 (ATCC^® CRL-2299^™)