Tissue engineering combines engineering principles and knowledge of life sciences to improve biological substituents. Three dimensional (3D) supporting structures, namely scaffolds obtained from biomaterials to mimic extracellular matrix (ECM) that provides suitable microenvironment for cell proliferation, migration and differentiation. In this study, poly (L-lactide-co-ε-caprolactone) (PLLCL) and collagen type I was used to fabricate scaffold by electrospinning method. In literature, collagen was often dissolved in toxic and harmful solvents that creates the major problem for cell culture applications. To overcome this problem “co-spinning” methodology is utilized for the formation of non-toxic collagen-based ECM mimetic scaffold. Collagen mixed with water-soluble carrier materials which is either polyvinylpyrrolidone (PVP) or polyvinyl alcohol (PVA) and co-electrospinning is carried out with PLLCL. Fabricated scaffolds were immersed into water to remove co-spinning agent; PVA or PVP, so only PLLCL/Collagen remained. PLLCL has homogeneous fibers in a diameter of 1.312 ± 0.22μm. The contact angle of PLLCL (136.6° ± 2.6) proved hydrophobic behavior of PLLCL material. The contact angle of the scaffold decreased up to 86.7° ± 0.1 confirming that hydrophobic behavior is decreased with the addition of collagen. Also, collagen-containing scaffolds were saturated at lower amount of protein than PLLCL, PLLCL/PVA and PLLCL/PVP scaffolds. Cytotoxicity analysis of scaffolds showed that PVA containing scaffolds had lower viability than PVP containing scaffolds; so most of the cell studies were carried out with PLLCL/ Collagen scaffolds fabricated by PVP cospinning. Cell proliferation on PLLCL/Collagen scaffolds found to be more favorable than PLLCL and PLLCL/PVP scaffolds.