Establishment of mouse gingival junctional epithelial cell line using a bioengineered tooth system

Area of Science:

• Oral biology
• Periodontal research
• Cell biology

Background:

• Junctional epithelium (JE) is crucial for periodontal tissue defense against bacteria.
• Understanding JE molecular mechanisms is limited by the lack of established cell lines.
• Existing research faces challenges due to the absence of in vitro models.

Purpose of the Study:

• To establish a novel immortalized junctional epithelium cell line for in vitro research.
• To characterize the gene expression and proliferative capacity of the new cell line.
• To facilitate future investigations into JE functions and interactions.

Main Methods:

• Isolation of green fluorescent protein (GFP)-expressing JE cells from bioengineered mouse teeth using flow cytometry.
• RNA sequencing to analyze gene expression profiles of isolated JE cells.
• Immortalization of JE cells using simian virus 40 large T antigen via lentiviral vectors.
• Assessment of cell proliferation and senescence (β-galactosidase assay).

Main Results:

• Successfully isolated GFP-positive JE cells with gene expression patterns similar to primary JE cells.
• Established an immortalized JE cell line maintaining proliferative activity for over 20 passages.
• Confirmed the absence of cellular senescence in the established cell line.
• The new cell line demonstrated consistent gene expression profiles compared to primary JE cells.

Conclusions:

• A novel, immortalized junctional epithelium cell line has been successfully established.
• This cell line provides a valuable tool for in vitro studies of periodontal tissue.
• The findings pave the way for deeper understanding of JE's role in preventing bacterial infections.