Improved Methods for the Stable Generation of Human Papillomavirus‐Driven Head and Neck Cancer Cell Lines

Cell lines are an essential tool in research, leading to new discoveries involving drug studies, prognosis, treatment outcomes, genomic abnormalities, and cellular pathway deviations. There is an ongoing need for new cell lines in cancer research. Cell lines are often initiated by using tissue explants or dissociating cells onto plastic; this proves ineffective with challenging cell lines. Here we report two improved protocols in establishing stable cell lines. The first takes the more classical approach but is paired with fluorescence-activated cell sorting (FACs). The second protocol involves coculturing with feeder cells, coupled with FACs. We demonstrate the use of these protocols in the establishment of 17 cell lines, including several cancer associated fibroblasts (CAFs) and a human papilloma virus (HPV)- positive HNSCC cell line.