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HBEC-5i
HBEC-5i
規(guī)格:
貨期:
編號:B238126
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
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產(chǎn)品名稱 HBEC-5i
商品貨號 B238126
Organism Homo sapiens, human
Tissue brain, cerebral cortex
Cell Type cerebral microvascular endothelium
Product Format frozen
Biosafety Level 2  [Cells contain SV-40 viral DNA sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease Normal
Applications Studies in cancer, tumor metastasis, endothelium function, cell trafficking, surface molecule interactions, microvascular changes of the brain caused by malaria
Shipping Information frozen
Storage Conditions LN2 vapor phase
Images Cell Micrograph for ATCC CRL-3245
Derivation HBEC-5i cells were derived from small fragments of human cerebral cortex obtained from patients who had died of various causes. These brains were devoid of any pathologic abnormalities. Isolation and purification procedures were performed and the cells were cultured. They were then transfected with plasmid containing SV40 large T antigen.
Antigen Expression The cells express von Willebrand's factor (vWF), VE-cadherin, occludin and cell adhesion molecules VCAM-1 and ICAM-1. (Wassmer S, et al. Platelets potentiate brain endothelial alterations induced by Plasmodium falciparum. Infect. Immun. 74(1): 645-653, 2006. PubMed: 16369021)
Comments HBEC-5i (ATCC® No. CRL-3245™) has been shown to retain many of the characteristics of endothelial cells. These cells express stable patterns of endothelial cell markers such as VE-cadherin, von Willebrand factor VIII, and peripheral occludin, in addition to CD54, CD40, and CSA, and they show an up-regulation of CD54 and CD106 upon TNF activation. HBEC-5i cells exhibit major features of cerebral endothelial cells, especially efficient tight-junction structures, as assessed by high transendothelial electric resistance and very low permeability to 70-kDa dextran.These immortalized cells, because they are a continuously renewable source of human cerebral microvascular endothelial cells, can be used as a replacement for primary human brain endothelial cells for many research studies.
Complete Growth Medium

The base medium for this cell line is DMEM:F12 (ATCC® 30-2006™). To make the complete growth medium, add the following components to the base medium: 

  • 40 µg/mL endothelial growth supplement (ECGS)
  • Fetal bovine serum (FBS; ATCC® 30-2020™) to a final concentration of 10%
Subculturing

Note: These cells are cultured on vessels coated with 0.1% Gelatin (ATCC® No. PCS-999-027). Use 1.0 mL of gelatin per 10 cm2 surface area. Incubate at 37.0°C for ≥ 45 minutes. Aspirate gelatin just prior to adding cells to vessel(s).



Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 
1. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC® No. 30-2200) or 0.25% Trypsin – 0.53mM EDTA (ATCC® No. 30-2101) solution to remove all traces of serum which contains trypsin inhibitor. 
2. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). 
     Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. 
3. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. 
4. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C.


Subcultivation Ratio: 1:4 to 1:10 is recommended.
Medium renewal: 2 to 3 times per week
Cryopreservation Complete Growth Medium, 92.5%; DMSO, 7.5%
Culture Conditions

Temperature:37oC

STR Profile TH01: 6,9.3
D5S818: 11,13
D13S317: 9,12
D7S820: 11
D16S539: 10,13
CSF1PO: 11,13
Amelogenin: X,Y
vWA: 14,18
TPOX:  8,11
Name of Depositor Kathryn Kellar, Ph.D.
Year of Origin February 1994
References

Wassmer S, et al. TGF-B1 released from activated platelets can induce TNF-stimulated human brain endothelium apoptosis: a new mechanism for microvascular lesion during cerebral malaria. J. Immunol. 176: 1180-1184, 2006. PubMed: 16394007

Claessens A, et al. A subset of group A-like var genes encodes the malaria parasite ligands for binding to human brain endothelial cells. Proc. Natl. Acad. Sci. USA 109(26): E1772-E1781, 2012. PubMed: 22619330

Wassmer S, et al. Platelets potentiate brain endothelial alterations induced by Plasmodium falciparum. Infect. Immun. 74(1): 645-653, 2006. PubMed: 16369021

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