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$Immortalized-Rat-Arachnoid-Cells---SV40\/hTERT$

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Immortalized-Rat-Arachnoid-Cells---SV40\/hTERT

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Immortalized-Rat-Arachnoid-Cells---SV40\/hTERT

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	Print Version
BioSafety Level	II
Organism	Sprawgue-Dawley Rats (21-23 day old female)
Source Organ	Brain
Growth Properties	Adherent
Morphology	Spindle-like appearance at low density, cobblestone appearance at confluency
Population Doubling	approximately 35.3 hr
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	vimentin, cytokeratin and desmoplakin
Applications	For research only. The Immortalized Rat Arachnoid Cells- SV40/hTERT is unique in its 1. Ability to form tight junctions as measured by TEER and radiochemical (ie. TEER value of 193 SEM ohm-cm^2 is comparable to 206 SEM ohm-cm^2 value of that of Caco-2 cells); 2. Increased population doubling time ; 3. Ability to frow on collagen sponges as well as two-dimensional membranes.
Immortalization Method	Serial passaging and transduction with recombinant lentiviruses carrying SV40 Large T antigen and hTERT gene
Description	The arachnoid cells are important in the maintenance of apical-basal polarity and the formation of tight junctions at the cerebrospinal fluid-blood barrier (CSF-blood barrier). The Immortalized Rat Arachnoid cells-SV40/hTERT derived from primary rat arachnoid cells maintained the ability to form intact epithelial monolayer and displayed normal immunohistochemical and morphological phenotypes. As such, the Immortalized Rat Arachnoid cell line is useful in physiological studies in modeling arachnoid granulations and cerebrospinal fluid flow. In addition, this cell line is also useful in pathological conditions such as hydrocephalus, arachnoid cysts, or other disorders.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow III medium available in ABM, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin(G255). Atmosphere: air, 95%; Carbon dioxide (CO₂), 5%.
Quality Control	1. Immunofluorescence and Western Blotting were used to confirm the presence of SV40 (82kDa) and hTERT (122kDa) expression. 2. Immortalized Rat Arachnoid Cells exhibit spindle-like appearance at low density before reaching confluency and “cobblestone” appearance after becoming fully confluent, which appeared to be identical to its primary counterparts. 3. Immunofluorescence and Western Blotting were used to confirm the presence of arachnoid cell markers such as vimentin, cytokeratin and desmoplakin.
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.