CD cell line has been established from a poorly differentiated squamous cell carcinoma of tongue. This is a first ever cell line established from an Indian gutka chewer. Cell line was characterized for morphology, ultrastructure, doubling time, expression of epithelial markers, DNA content, karyotyping, STR markers, p53 mutations, HPV status, and tumorigenicity in SCID mice with all-trans-retinoic acid and cisplatin. The epithelial phenotype of the cell line was confirmed with surface markers and ultrastructure. The cell line is hyperploid with chromosomal alterations like gain of chromosomes 8q and 11q. CD cell line shows a unique pattern on STR genotyping and carries a missense mutation R273C inTP53. It does not show genomic integration of HPV. The cells are nontumorigenic to SCID mice and show growth inhibition upon treatment with cisplatin, and all-trans-retinoic acid. This cell line may be useful as an in vitro tool to understand the molecular changes associated with oral cancers.