Synthetic and natural polymer association is a promising tool in tissue engineering. The aim of this study was to compare five methodologies for producing hybrid scaffolds for cell culture using poly-l-lactide (PLLA) and collagen: functionalization of PLLA electrospun by (1) dialkylamine and collagen immobilization with glutaraldehyde and by (2) hydrolysis and collagen immobilization with carbodiimide chemistry; (3) co-electrospinning of PLLA/chloroform and collagen/hexafluoropropanol (HFP) solutions; (4) co-electrospinning of PLLA/chloroform and collagen/acetic acid solutions and (5) electrospinning of a co-solution of PLLA and collagen using HFP. These materials were evaluated based on their morphology, mechanical properties, ability to induce cell proliferation and alkaline phosphatase activity upon submission of mesenchymal stem cells to basal or osteoblastic differentiation medium (ODM). Methods (1) and (2) resulted in a decrease in mechanical properties, whereas methods (3), (4) and (5) resulted in materials of higher tensile strength and osteogenic differentiation. Materials yielded by methods (2), (3) and (5) promoted osteoinduction even in the absence of ODM. The results indicate that the scaffold based on the PLLA/collagen blend exhibited optimal mechanical properties and the highest capacity for osteodifferentiation and was the best choice for collagen incorporation into PLLA in bone repair applications.