C8919 can be used for flask coating in cell culture; however, coating protocols should be optimized for the SCC066 cell line, since explicit data for this cell line is not provided. For SCC066, rat tail collagen 08-115 is recommended; please follow the protocol that is available on the SCC066 product page.
SCC066
Blood-Brain Barrier hCMEC/D3 Cell Line
Human
Synonym(s):
Human BBB cell line, Human blood-brain barrier cell line, Human cerebral endothelial cell line, CMEC/D3 cell Line, hCMEC/D3 cell line
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About This Item
UNSPSC Code:
41106514
NACRES:
NA.81
eCl@ss:
32011203
Biological source:
human
Pricing and availability is not currently available.
Product Name
Blood-Brain Barrier hCMEC/D3 Cell Line, The hCMEC/D3 BBB cell line has been extensively characterized for brain endothelial phenotype and is a model of human blood-brain barrier (BBB) function.
biological source
human
Quality Segment
technique(s)
cell culture | mammalian: suitable, drug transporter assay: suitable
shipped in
liquid nitrogen
storage temp.
-140 to -196°C
General description
The blood–brain barrier (BBB) is a highly selective permeability barrier that separates the circulating blood from the brain extracellular fluid in the central nervous system. The blood–brain barrier is formed by capillary endothelial cells, which are connected by tight junctions with an extremely high electrical resistivity. The blood–brain barrier allows the passage of water, some gases, and lipid soluble molecules by passive diffusion, as well as the selective transport of molecules such as glucose and amino acids that are crucial to neural function. On the other hand, the blood–brain barrier may prevent the entry of lipophilic, potential neurotoxins by way of an active transport mechanism mediated by P-glycoprotein. Astrocytes are necessary to create the blood–brain barrier. The blood–brain barrier (BBB) prevents entry into the brain of most drugs from the blood. The presence of the BBB makes difficult the development of new treatments of brain diseases, or new radiopharmaceuticals for neuroimaging of brain.
The hCMEC/D3 cell line was derived from human temporal lobe microvessels isolated from tissue excised during surgery for control of epilepsy. The primary isolate was enriched in cerebral endothelial cells (CECs). In the first passage, cells were sequentially immortalized by lentiviral vector transduction with the catalytic subunit of human telomerase (hTERT) and SV40 large T antigen, following which CEC were selectively isolated by limited dilution cloning, and clones were extensively characterized for brain endothelial phenotype. This brain microvascular endothelial cell line represents one such model of the human BBB that can be easily grown and is amenable to cellular and molecular studies on pathological and drug transport mechanisms with relevance to the central nervous system (CNS).
Reference:
Couraud PO, et al. (2008) The human brain endothelial cell line hCMEC/D3 as a human blood‐brain barrier model for drug transport studies. Fluids Barriers CNS. 2013 Mar 26;10(1):16.
Reference:
Couraud PO, et al. (2008) The human brain endothelial cell line hCMEC/D3 as a human blood‐brain barrier model for drug transport studies. Fluids Barriers CNS. 2013 Mar 26;10(1):16.
Application
Research Category
Neuroscience
Neuroscience
Research Sub Category
Neurodegenerative Diseases
Neurodegenerative Diseases
Subject to local law, this product is intended to be sold for internal in vitro research use only subject to terms and conditions found here: www.sigmaaldrich.com/restrictedcelluse. This product may not be: re-engineered or copied; used to make derivatives, modifications or functional equivalents; used to obtain patents or other IP claiming use of the product; used to develop, test, or manufacturer a commercial product; used as a component in a commercial product; resold or licensed; used in any clinical applications or trials; or used in humans. A license or limited commercial use agreement is required for use by any for-profit entity, use in services, and use in sponsored academic research. For information regarding any such use, please contact [email protected].
The hCMEC/D3 BBB cell line has been extensively characterized for brain endothelial phenotype and is a model of human blood-brain barrier (BBB) function.
Biochem/physiol Actions
Endothelial Cells
Preparation Note
Blood-Brain Barrier hCMEC/D3 cells should be stored in liquid nitrogen. The cells can be passage for at least 10 passages without significantly affecting the cell marker expression and functionality.
Analysis Note
• Each vial contains ≥ 1X10^6 viable cells.
• Cells are tested by PCR and are negative for Hepatitis A, B, C, HPV, Herpes and HIV-1 & 2 viruses.
• Cells are negative for mycoplasma contamination.
• Cells are tested by PCR and are negative for Hepatitis A, B, C, HPV, Herpes and HIV-1 & 2 viruses.
• Cells are negative for mycoplasma contamination.
Disclaimer
RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges {HCompany} to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges {HCompany} to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
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This Item | |||
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| technique(s) cell culture | mammalian: suitable, drug transporter assay: suitable | technique(s) cell culture | stem cell: suitable | technique(s) cell based assay: suitable | technique(s) cell based assay: suitable, cell culture | mammalian: suitable, cell migration: suitable |
| biological source human | biological source rat | biological source mouse | biological source human |
| shipped in liquid nitrogen | shipped in liquid nitrogen | shipped in dry ice | shipped in liquid nitrogen |
| storage temp. -140 to -196°C | storage temp. -140 to -196°C | storage temp. -140 to -196°C | storage temp. −196°C |
| Quality Level 100 | Quality Level 100 | Quality Level 100 | Quality Level 100 |
Storage Class
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| SCC066 | 04055977276138 |
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Could I use C8919-20ML to coat my flask for this cell line?
1 answer-
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Do they grow on Transwell? where can we find the media to grow it?
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According to literature, this cell line can be grown on Transwell. Cells are cultured in EndoGRO™-MV Complete Media Kit supplemented with 1 ng/mL FGF-2.
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Could you tell me if the cells were originally isolated from a male or a female donor?
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Human brain tissue was obtained from an adult female donor.
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What is the passage number we should expect for the vial?
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The passage number is dependent on the lot. The cells can be passaged for at least 10 passages without significantly affecting cell marker expression and functionality.
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Hi, regarding cell line hCMEC/ D3 (SCC066): Since cell line was immortalized by transfection using lentivirus, I would like to know if you have information on the on the biosafety of the cell line regarding potential expression of virus/ viral products?
1 answer-
The Biosafety Level for this product has been classified as Risk Group 2 based on both US and EU classifications
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Hi, I hope you are well. I was wondering which generation of lentivirus was used to produce this cell line? Could you please provide more information regarding the safety of this cell line? Thank you, Ecem
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These cells are classified as GMO RSG-1 in Europe - this was determined by the German central committee on biological safety because of the following:
"transduced cells were tested for any production of retroviral particles, using a sensitive enzyme assay for p24 (not shown) in the first and second passage, and both cells and conditioned media were negative."
Please see the originating paper for this statement by the authors.The Vector:
Lentivirus: (name and class not known).The transgene/s:
The cells are immortalized by transduction with the transgenes for the catalytic subunit of human telomerase (hTERT) and SV40 large T antigen.Please also see the Cellosaurus entry: https://www.cellosaurus.org/CVCL_U985
Safe handling:
These cells should be used at a BSL-2 (not necessarily due to GMO) but because whilst all efforts are taken to screen for common adventitious agents (e.g. Hepatitis A, B, C, HPV, Herpes and HIV-1 & 2 viruses, mycopasma) they can never cover all diseases/infections.Helpful?
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