In this study, we successfully produced the chitosan microfibers using the proposed various angles of microfluidic chip, which was also been simulated. By controlling the core and sheath flow rates, we were able to generate laminar flow of different diameters from 15 μm to 40 μm. And the diameter of chitosan microfiber was measured from 20 μm to 50 μm. The microchannel of angle 30° could produce chitosan laminar flow of a smaller diameter than the angle 60° and angle 45° at the fixed flow rates. Finally, the chitosan microfiber was chosen as scaffold and the schwann cell and fibroblast cell with chitosan microfibers were used for cell culture to test effect in tissue engineering application.