Collagen is an extracellular matrix protein with great importance in biomedical application. The search for collagen from various sources is intensified especially from marine source. This study was carried out to extract collagen from a Malaysian local fresh water fish, Clarias batrachus and characterized its biomedical potential In vitro. Collagen was extracted from C. batrachus skin using acetic acid method and identified using SDS-PAGE. MTT assay was performed to determine the effect of coated collagen on cell adhesion and proliferation of L929 skin fibroblast cells. Additionally, scratch assay was performed to examine the effect of collagen coating on fibroblast cell migration. Result showed that collagen extracted from C. batrachus was made up of collagen type I, which consists of two α chains (α-1 and α-2) and β chain. At 100 µg/cm 2 density, collagen coating significantly increased fibroblast cell adhesion, proliferation and migration compared to negative control (p< 0.05). As a conclusion, collagen extracted from C. batrachus increased cell adhesion, proliferation and migration of fibroblast cells and has potential to be used as an alternative source of collagen.