Background and Objectives The Cholesterol granuloma is often observed in middle ear surgeries. In the clinic, it can be found in a normal ear, but also as a recurrent serous otitis media in non-responding medical therapy. The purpose of this study is to analyze the clinical characteristics of cholesterol granuloma with or without cholesteatoma. Subjects and Method We performed a retrospective study of 40 patients who underwent tympanomastoidectomy between January, 2000 and January, 2009 for cholesterol granuloma. We divided patients into a non-cholesteatoma group and a cholesteatoma group and observed the following clinical manifestations: initial symptom findings, ear drum findings, preoperative audiogram, postoperative audiogram, operative technique and operative findings. Results Cholesterol granuloma was found in 10.2% of 393 cases (40 cases) who underwent chronic otitis media surgery. Of these, 5.3% (21 cases) was diagnosed with non-cholesteatoma group and 4.8% (19 cases) was diagnosed in the cholesteatoma group. Chief complaints were the difficulty of hearing in the non-cholesteatoma group and otorrhea in the cholesteatoma group. Ear drums were nearly retracted and perforated in both groups, although the cholesteatoma group had more damaged ear drum. Distinctive features of hearing were the conductive type with a moderate degree and ossicular destruction showed missing or eroded incus. Major sites of cholesterol granuloma were antrum, air cells and epitympanum. Conclusion Although ear drum was relatively preserved in the non-cholesteatoma group of cholesterol granuloma, the hearing level and the degree of ossicular destruction were not so much different between the two groups. Therefore, a complete removal of cholesterol granuloma and ossiculoplasty should be considered for better hearing restoration.