A new bovine cell line was developed from tumor biopsy material of rectum obtained from clinical case of 7 years old cattle with tumor mass obliterating the rectal opening. Histopathology of tumor revealed scattered stellate cells arranged singly or in clusters in loose mucinous ground substance, simulating myxoma. The cells obtained from tumor mass have been cultured for more than 36 months in DMEM supplemented with 10% fetal bovine serum (FBS). The population doubling time of this cell line was about 20.64 h. The cytogenetic analysis revealed several chromosomal abnormalities with bizarre karyotype. The origin of the cell line was confirmed by PCR amplification of 1086 bp fragment of 16s rRNA using bovine species specific primers. The new cell line would act as in vitro model to study many aspect of cancer biology such as tumor development, differentiation and therapeutics regimen to combat cancer.