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. Immortalization Method: Serial passaging and transduction with recombinant retroviruses carrying hTERT geneBioSafety Level: IISpecies: HumanSource Organ: GingivaGrowth Properties: AdherentMorphology: Spindle shapedPopulation Doubling: 50-60 hoursSeeding Density: 7,000-10,000 cells/cm10²; Recommended split ratio is 1:2 to 1:3Markers: Vimentin and actinDonor Age: N/ADonor Gender: N/ADonor Ethnicity: N/APropagation: The base medium for this cell line is Prigrow III medium available at 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 Solution (G255) to a final concentration of 1% Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.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. Shipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C