The purpose of this study is to make use of trans10,cis12 CLA (t-CLA) that has potential for proliferation and differentiation to form adipocyte on the collagen-coated surface. Results provided evidences of good adhesion, growth, viability, and differentiation of adipocyte on collagen-coated surface compared with non-coated surface. Also, the results showed that mouse 3T3-L1 preadipocyte can be successfully and reproducibly cultured on the collagen-coated surface, and the adipocyte precursor cells placed on the collagen-coated surface are able to undergo full maturation into adipocytes in the control cells. Moreover, glycerol-3-phosphate dehydrogenase (GPDH) activity in 3T3-L1 preadipocyte cultured on collagen-coated surface with t-CLA was higher than that on polystyrene (PS) surface due to higher cell adhesion and cell viability. These results suggest that collagen coating may provide a promising approach to develop a new adipocyte replacement strategy using CLA.