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微信小章| 产品名称： | HBEC-5i |
|---|---|
| 商品货号： | TS138794 |
| Organism： | Homo sapiens, human |
| Tissue： | brain, cerebral cortex |
| Cell Type： | cerebral microvascular endothelium |
| Product Format： | frozen |
| Biosafety Level： | 2 xa0Cells 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： | LN |
| Images： |  |
| 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 Willebrands 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™)xa0has 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:xa0
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| Subculturing： | Note: These cells are cultured on vessels coated with 0.1% Gelatin (ATCC®xa0No. 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.xa0 1. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos phosphate-buffered saline (D-PBS) (ATCC®xa0No. 30-2200) or 0.25% Trypsin – 0.53mM EDTA (ATCC®xa0No. 30-2101) solution to remove all traces of serum which contains trypsin inhibitor.xa0 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).xa0 xa0 xa0 xa0Note: 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.xa0 3. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.xa0 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: xa08,11 |
| 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 |