This study provides original evidence demonstrating that CAF-secreted factors sustain and robustly enhance stemness in HNSCC-derived cell lines, thereby increasing anchorage-independent growth, tumorsphere formation, and expression of various CSC markers, such as NANOG, SOX2, OCT4, ALDH1, CD133, CD44, and NOTCH1, in the absence of serum or any other supplements. Here, NANOG is linked to head and neck squamous cell carcinoma.