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Immortalized-Human-Gingival-Fibroblasts--hTERT

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Immortalized-Human-Gingival-Fibroblasts--hTERT

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Immortalized-Human-Gingival-Fibroblasts--hTERT

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Gingiva
Growth Properties	Adherent
Morphology	spindle-shaped
Population Doubling	50h
Recommended Seeding Density	5,000-7,000 cells per cm^2
Markers	Vimentin and actin
Applications	For research only. The Immortalized Human Gingival Fibroblast -hTERT is unique that 1. The cells retained similar cytotoxicity response pattern to their primary counterparts (in response to phenol, gold, titanium and amalgam); 2. The cells retained identical phenotype to that of their primary counterparts; 3. The cells have a diploid status, showing genomic stability.
Immortalization Method	Serial passaging and transduction with recombinant lentiviruses carrying hTERT gene
Description	Gingival fibroblasts play an important role in oral wound healing and are the major constituents of the gingival connective tissue. These cells are important in testing the “biocompatibility”, or toxicity of new dental materials. In the past, toxicity tests in vitro have been performed with non-human and human malignant cell lines such as L929, CCL163, CCL171 and SaOS2. However, due to the malignancy of the described cell lines, the results may or may not be clinical relevant. The Immortalized Human Gingival Fibroblasts- hTERT have the same morphological cellular response in biocompatibility evaluation to primary human gingival fibroblasts and can thus be used to evaluate toxicity of dental materials.
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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, RT-PCR and Western Blotting were used to confirm the presence of hTERT expression (PCR primers for hTERT: 5’-CGGAAGAGTGTCTGGAGCAA-3’ forward, and 5’-GGATGAAGCGGAGTCTGGA-3’ reverse). 2. MTT assay was used to confirm the proliferative potential of the immortalized cells over the 70th passage.
Reference	Kim, J., et al. (2012). Immortalized Gingival Fibroblasts as a Cytotoxicity Test Model for Dental Materials. J. Mater Sci: Mater Med 23:753-762.
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.