The current study aims at bacterial and fungal etiology of Cholesteatoma in patients attending tertiary care hospital. Cholesteatoma is an abnormal, noncancerous skin growth that can develop in the middle section of ear, behind the eardrum. It is a complication of chronic suppurative otitis media. With the informed consent, 70 patients with Cholesteatoma were included; 42 (60%) were male and 28 (40%) were female, and the mean age was 11-50 years.