cell biology
atcc® number: crl-2299™    
designations: bend.3
depositors:  ec butcher
biosafety level: 1
shipped: frozen
medium & serum: seepropagation
growth properties: adherent
organism: musmusculus
morphology: endothelial
source: organ: brain 
strain: balb/c 
tissue: cerebral cortex 
disease: endothelioma 
cell type: endothelialpolyoma middle tantigen transformed
cellular products: von willebrandfactor
permits/forms: in addition tothe mta mentioned above, other atcc and/orregulatory permits may be required for the transfer of thisatcc material. anyone purchasing atcc material is ultimatelyresponsible for obtaining the permits. please clickhere for information regarding the specific requirements forshipment to your location.
restrictions: the bend.3 cell line isprovided for research use only under the following terms:1. bend.3cells, or products derived from them, must not be sold or used forcommercial purposes. nor can the cells be distributed to thirdparties for purpose of sale, or producing for sale, bend.3 cells ortheir products. commercial interests are the exclusive property ofthe max planck society. 2. any proposed commercial use of bend.3cells, or products derived from them, must first be negotiated withdr. 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:. in all papers reporting any use ofbend.3 cells, or products derived from them, a direct referencewill be made to the original publication (montesano, et al. cell62:435-445, 1990).
applications: the cells weretransformed by infection with the ntkmt retrovirus vector thatexpresses polyomavirus middle t antigen. 
the endothelial nature of these cells was confirmed by the observedexpression of von willebrand factor and uptake of fluorescentlylabeled low density lipoprotein (ldl).
antigen expression: icam-1 +; vcam-1 +;madcam-1 +
age: 6 weeks
comments: the cells weretransformed by infection with the ntkmt retrovirus vector thatexpresses polyomavirus middle t antigen. 
the endothelial nature of these cells was confirmed by the observedexpression of von willebrand factor and uptake of fluorescentlylabeled low density lipoprotein (ldl). 
the expression of peyer's patch high endothelial receptor forlymphocytes, the mucosal vascular addressin (madcam-1) ande-selectin can be induced on bend.3 cells by cytokines andlipopolysaccharide (lps). 
this induction by tumor necrosis factor alpha (tnf alpha),interleukin 1 (il-1) or lps is concentration and timedependent. 
madcam-1 is expressed on the surface of unstimulated bend.3 cellsat early passages but not at passages greater than 30. 
intracellular adhesion molecule 1 (icam-1) is constitutivelyexpressed on the cells, and expression is increased by treatmentwith lps, il-1 and tnf alpha. 
vascular cell adhesion molecule 1 (vcam-1) is constitutivelyexpressed on the cells at early passages but not at passages over30. 
p-selectin can be induced on bend.3 cells by tumor necrosis factoralpha (tnf alpha) at both early and late passages but expression isgreater at passages over 30.
propagation: atcc completegrowth medium: the base medium for this cell line isatcc-formulated dulbecco's modified eagle's medium, catalog no.30-2002. to make the complete growth medium, add the followingcomponents to the base medium: fetal bovine serum to a finalconcentration of 10%.
temperature: 37.0°c 
atmosphere: air, 95%; carbon dioxide (co2),5%
subculturing: protocol: removeand discard culture medium. briefly rinse the cell layer with 0.25%(w/v) trypsin-0.03% (w/v) edta solution to remove all traces ofserum which contains trypsin inhibitor.add 1.0 to 2.0 ml oftrypsin-edta solution to flask and observe cells under an invertedmicroscope until cell layer just begins to detach. add 6.0 to 8.0ml of complete growth medium and aspirate cells by gentlypipetting. add appropriate aliquots of the cell suspension to newculture vessels. 
subcultivation ratio: a subcultivation ratioof 1:6 to 1:10 is recommended 
medium renewal: every 2 to 3 days
preservation: freezemedium: complete growth medium, 95%; dmso,5% 
storage temperature: liquid nitrogen vaportemperature
related products: recommended medium(without the additional supplements or serum described under atccmedium): atcc 30-2002
recommended serum: atcc 30-2020
references: 39039:montesano r, et al. increased proteolytic activity is responsiblefor the aberrant morphogenetic behavior of endothelial cellsexpressing the middle t oncogene. cell 62: 435-445, 1990.pubmed: 237923739040: sikorski ee,et al. the peyer's patch high endothelial receptor for lymphocytes,the mucosal vascular addressin, is induced on a murine endothelialcell line by tumor necrosis factor-alpha and il-1. j. immunol. 151:5239-5250, 1993. pubmed: 769380739041: williams rl, et al. embryonic lethalities andendothelial tumors in chimeric mice expressing polyoma virus middlet oncogene. cell 52: 121-131, 1988.pubmed: 3345558

---

上海迪奥生物科技有限公司
陈
13816767942
斜土路1175号景泰大厦1508室