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Immortalized Human Gingival Keratinocytes (Gie-No3B11)

Cat.No.: CSC-I9236L

Species: homo sapiens

Source: Buccal Gingiva

Morphology: Polygonal

Culture Properties: Adherent

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Cat.No.
CSC-I9236L
Description
Immortalized Human Gingival Keratinocytes-hTERT were developed from human tissues transduced with a lentiviral expression vector containing the hTERT gene. The cell line was continuously cultured for more than 20 passages without showing signs of growth retardation or replicative senescence.
Species
homo sapiens
Source
Buccal Gingiva
Recommended Medium
SuperCult® Immortalized Human Gingival Keratinocyte Medium (Cat No.: CM-I9236L)
Freezing Medium
Complete medium supplemented with 10% (v/v) DMSO
Culture Properties
Adherent
Morphology
Polygonal
Immortalization Method
Human telomerase reverse transcriptase (hTERT)
Application
For Research Use Only
Growth Properties
Cells are cultured as a monolayer at 37°C in a humidified atmosphere with 5% CO2.
Storage
Directly and immediately transfer cells from dry ice to liquid nitrogen upon receiving and keep the cells in liquid nitrogen until cell culture needed for experiments.

Note: Never can cells be kept at -20 °C.
Shipping
Dry Ice.
Recommended Products
CIK-HT013 HT® Lenti-hTERT Immortalization Kit
CIK-HT003 HT® Lenti-SV40T Immortalization Kit
Quality Control
Real Time PCR was used to quantify hTERT gene expression in immortalized cell line. Free from contaminations (bacteria incl. mycoplasma, fungi, HIV, HAV, HBV, HCV, Parvo-B19) and cross-contaminations.
BioSafety Level
II
Citation Guidance
If you use this products in your scientific publication, it should be cited in the publication as: Creative Bioarray cat no. If your paper has been published, please click here to submit the PubMed ID of your paper to get a coupon.

Primary gingival keratinocytes extracted from human oral gingival tissue underwent immortalization to create the Gie-No3B11 cell line. The oral mucosa contains gingival tissue which is located between the teeth and the alveolar bone. Gingival tissue acts as a protective barrier that keeps periodontal tissue safe from bacteria and harmful substances. The Gie-No3B11 cell line displays morphological characteristics which are identical to those seen in primary gingival keratinocytes. The cells generate cytokeratins such as cytokeratin 2 alongside cytokeratin 17 and cytokeratin 19 and exhibit standard polygonal or irregular keratinocyte shapes. Furthermore, these cells show the capability to develop dense multilayer structures.

Scientists use the Gie-No3B11 cell line as a standard model to study both periodontal disease development and oral mucosal cancer progression. They can also be co-cultured with other cells (such as fibroblasts) to construct three-dimensional tissue equivalents, which are used to study tissue repair and regeneration processes.

Metabolic Activity of Gingival Epithelial Cells in Response to Core-Multishell Nanocarriers

Drug delivery systems target the oral mucosa because it provides easy access and avoids hepatic first-pass metabolism. The bioavailability of drugs used in local therapy for oral diseases decreases because of the mucosal barrier and the diluting effects of saliva and crevicular fluid. To address this, Jager et al. characterized core-multishell (CMS) nanocarriers for their potential use in oral mucosal drug delivery.

To evaluate potential cytotoxic effects of the CMS nanocarrier, we performed MTT assays to monitor the metabolic activity (cell survival) of cells after CNS nanocarrier application (Fig. 1). They used the immortalized gingiva keratinocytes Gie-No3B11 due to an intended topical use of the particles at the oral mucosa. The application of the unloaded CMS nanocarrier did not affect viability of the cells at concentrations ranging from 1 to 50 μg/mL after 24 hours. The higher concentrations 100 and 500 μg/mL slightly decreased the metabolic activity when compared to control epithelial cells (Fig. 1A). After 48 hours, CMS nanocarriers did not influence epithelial cell survival at any concentration tested (Fig. 1B). After 72 hours, the metabolic activity was unaffected at CMS concentrations between 1 and 100 μg/mL, and a reduced metabolic activity was found for the highest concentration (500 μg/mL) applied compared to control cells (Fig. 1C).

The metabolic activity of gingival epithelial cells was assessed using the MTT assay after exposure to varying concentrations of the CMS nanocarrier for 24 hours (A), 48 hours (B), or 72 hours (C).Fig. 1. Metabolic activity in gingival epithelial cells was monitored when exposed to different concentrations the CMS nanocarrier for 24 (A), 48 (B) or 72 hours (C) using the MTT assay (Jager J, Obst K, et al., 2018).

Glycation of Host Proteins Increases Pathogenic Potential of Porphyromonas gingivalis

The connection between periodontitis and diabetes exists because diabetes raises periodontitis risk through increased inflammatory responses and protein glycation. The non-enzymatic process of glucose binding to proteins known as glycation occurs frequently in diabetic individuals and can modify protein functions. Śmiga et al. aim to determine if glycated proteins enhance the growth and pathogenic potential of Porphyromonas gingivalis, a key periodontal pathogen.

They found that glycation increases the ability of P. gingivalis to acquire heme from hemoglobin, mostly due to heme sequestration by the HmuY hemophore-like protein. Based on their results, they hypothesize that differences in hemoglobin degradation and heme release between glycated and un-glycated hemoglobin may not significantly enhance P. gingivalis growth in laboratory liquid culture media. P. gingivalis-mediated diseases involve the bacterium's ability to infect host cells and evade immune defenses. We examined how P. gingivalis infected keratinocytes cultured with hemoglobin as the sole heme source. Glycated hemoglobin slightly increased P. gingivalis infection ability, mainly through enhanced bacterial attachment rather than invasion (Fig. 2). The evidence indicates that glycation could permit P. gingivalis survival and deeper penetration into the epithelium through intercellular pathways. Statistical analysis revealed no significant differences between cultures containing glycated hemoglobin and those containing un-glycated hemoglobin. The simple keratinocyte culture model used for this test likely fails to completely simulate real-life in vivo conditions. Research with a three-dimensional gingival model featuring collagen microfibers revealed comparable results.

Formation of the HmuY-Fe(III)heme complex from hemoglobin was compared.Fig. 2. Comparison of HmuY-Fe(III)heme complex formation from hemoglobin (Śmiga M, Smalley JW, et al., 2021).

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